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Histology Objective Lenses

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Neil Anthony

Histology Objective Lenses

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Hi all, I hope the scopes treat you well.

I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.

Not having much histology experience, I have to ask:
If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?

Thanks in advance for your time.

Neil

________________________________

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prohibited.

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Neil Anthony

Re: Histology Objective Lenses

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And for completeness, the below email is not fraudulent, and I think the auto system has a false positive. Unless of course you tell me your trade secrets about objective lenses that I can sell for big money, then things might be different ;)

Neil

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Anthony, Neil
Sent: Friday, June 30, 2017 1:10 PM
To: [hidden email]
Subject: Histology Objective Lenses

[This sender failed our fraud detection checks and may not be who they appear to be. Learn about spoofing at http://aka.ms/LearnAboutSpoofing]

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Hi all, I hope the scopes treat you well.

I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.

Not having much histology experience, I have to ask:
If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?

Thanks in advance for your time.

Neil

________________________________

This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited.

If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments).

TSwayne

Re: Histology Objective Lenses

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Hi Neil,

Two types of lenses we use all the time for histology are:
1) very low-mag (like 2.5x and 5x) for documenting large areas, and
2) “high-dry” (meaning high NA, no oil) like 10x/0.5, 20x/0.8 or 40x/0.8.
“Plan” (flat-field corrected) lenses give you good correction across those wide fields.

We don’t use oil on histology samples much, but if you did have the budget for an oil lens, 60x should be enough for most users and high-pixel-count cameras.

If you will have users walking in with cell culture plates or chamber slides with living cells, you might also want a long-working-distance lens… probably 20x or maybe 40x.
I would definitely get a phase ring on that lens (some vendors have phase rings in the lens, others not). A correction collar for substrate thickness will give you a sharper image on a wider variety of samples.

Hope this helps, and happy shopping!
Theresa

On Jun 30, 2017, at 2:19 PM, Anthony, Neil <[hidden email]<mailto:[hidden email]>> wrote:

Hi all, I hope the scopes treat you well.

I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.

Not having much histology experience, I have to ask:
If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?

Thanks in advance for your time.

Neil

------------------------------------
Theresa Swayne, Ph.D.
Manager
Confocal and Specialized Microscopy Shared Resource<http://hiccc.columbia.edu/research/sharedresources/confocal>

Herbert Irving Comprehensive Cancer Center
Columbia University Medical Center
1130 St. Nicholas Ave., Room 222A
New York, NY 10032
Phone: 212-851-4613
[hidden email]<mailto:[hidden email]>

Martin Wessendorf-2

Re: Histology Objective Lenses

In reply to this post by Neil Anthony

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Dear Neil--

First, I think that the "fraudulent email" alert must be local to
Emory--I've never seen it on any posting to the list, including yours!

Regarding objectives--there could be a lot of answers to this question,
depending in part on your budget and on your anticipated uses. I have a
turret with 6 objectives, four of which I use daily (4x, 10x, 20x, and
40x dry)--I'd have a hard time living without any of those. (I use my
2x and 60x oil lenses less often.) However, magnification doesn't tell
the full story--we also need to talk about objective type
(e.g.,acromats, fluorites, planapochromats, etc.), numerical aperture,
transmittance at different wavelengths, etc. What you want will depend
on what you will be looking at and recording.

--Could you talk to your dealer and see if they could lend you a full
set for an extended period, from which you could pick the ones that you
like?

Good luck!

Martin

On 6/30/2017 1:19 PM, Anthony, Neil wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> And for completeness, the below email is not fraudulent, and I think the auto system has a false positive. Unless of course you tell me your trade secrets about objective lenses that I can sell for big money, then things might be different ;)
>
> Neil
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Anthony, Neil
> Sent: Friday, June 30, 2017 1:10 PM
> To: [hidden email]
> Subject: Histology Objective Lenses
>
> [This sender failed our fraud detection checks and may not be who they appear to be. Learn about spoofing at http://aka.ms/LearnAboutSpoofing]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi all, I hope the scopes treat you well.
>
> I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.
>
> Not having much histology experience, I have to ask:
> If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?
>
> Thanks in advance for your time.
>
> Neil
>
>
>
>
>
>
>
>
>
> ________________________________
>
> This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited.
>
> If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments).

mcammer

Re: Histology Objective Lenses

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Post images on http://www.imgur.com and include the link in your posting.
*****

I have found that most people get inexpensive lenses which is fine for scanning by eye when looking at the center of the field, but if you want to take pictures and make mosaics, go for planapochromat so that all colors are in focus and when you do tiling, the edges of the fields are linear.

=*===========================================================*=
Michael Cammer, DART Microscopy Laboratory, NYU Langone Medical Center
Cell: 914-309-3270 (this is for calling, not texting) Office: Skirball 2nd Floor main office, back right
http://ocs.med.nyu.edu/microscopy & http://microscopynotes.com/

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Martin Wessendorf
Sent: Friday, June 30, 2017 2:57 PM
To: [hidden email]
Subject: Re: Histology Objective Lenses

Dear Neil--

First, I think that the "fraudulent email" alert must be local to Emory--I've never seen it on any posting to the list, including yours!

Regarding objectives--there could be a lot of answers to this question, depending in part on your budget and on your anticipated uses. I have a turret with 6 objectives, four of which I use daily (4x, 10x, 20x, and 40x dry)--I'd have a hard time living without any of those. (I use my 2x and 60x oil lenses less often.) However, magnification doesn't tell the full story--we also need to talk about objective type (e.g.,acromats, fluorites, planapochromats, etc.), numerical aperture, transmittance at different wavelengths, etc. What you want will depend on what you will be looking at and recording.

--Could you talk to your dealer and see if they could lend you a full set for an extended period, from which you could pick the ones that you like?

Good luck!

Martin

On 6/30/2017 1:19 PM, Anthony, Neil wrote:

> *****
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> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DQICaQ&c=j5oPpO0eBH1iio48DtsedbOB
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> wEc8jjjrMeD9SR4lkd7JOS3aNGGb4OHv2tdXepSZY&s=unk8s8IQUYA_xAey7ycnWVTrEv
> jgv2xETE24LtNGQLU&e= Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DQICaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=JgwEc8jjjrMeD9SR4lkd7JOS3aNGGb4OHv2tdXepSZY&s=vkaXMcoLi3mZU4Pw1nFbtSyzH8OLq8aanLVsIyOxvcQ&e= and include the link in your posting.
> *****
>
> And for completeness, the below email is not fraudulent, and I think
> the auto system has a false positive. Unless of course you tell me
> your trade secrets about objective lenses that I can sell for big
> money, then things might be different ;)
>
> Neil
>
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of Anthony, Neil
> Sent: Friday, June 30, 2017 1:10 PM
> To: [hidden email]
> Subject: Histology Objective Lenses
>
> [This sender failed our fraud detection checks and may not be who they
> appear to be. Learn about spoofing at
> https://urldefense.proofpoint.com/v2/url?u=http-3A__aka.ms_LearnAboutS
> poofing&d=DQICaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05
> LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=JgwEc8jjjrMeD9SR4lkd7JOS3aNGG
> b4OHv2tdXepSZY&s=HDvoq9eJq_Pkd4zXWmydwuCKH2BPJLEowI8yeIs4JSE&e= ]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DQICaQ&c=j5oPpO0eBH1iio48DtsedbOB
> Gmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=Jg
> wEc8jjjrMeD9SR4lkd7JOS3aNGGb4OHv2tdXepSZY&s=unk8s8IQUYA_xAey7ycnWVTrEv
> jgv2xETE24LtNGQLU&e= Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DQICaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=JgwEc8jjjrMeD9SR4lkd7JOS3aNGGb4OHv2tdXepSZY&s=vkaXMcoLi3mZU4Pw1nFbtSyzH8OLq8aanLVsIyOxvcQ&e= and include the link in your posting.
> *****
>
> Hi all, I hope the scopes treat you well.
>
> I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.
>
> Not having much histology experience, I have to ask:
> If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?
>
> Thanks in advance for your time.
>
> Neil
>
>
>
>
>
>
>
>
>
> ________________________________
>
> This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited.
>
> If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments).

--
Martin Wessendorf, Ph.D. office: (612) 626-0145
Assoc Prof, Dept Neuroscience lab: (612) 624-2991
University of Minnesota Preferred FAX: (612) 624-8118
6-145 Jackson Hall, 321 Church St. SE Dept Fax: (612) 626-5009
Minneapolis, MN 55455 e-mail: [hidden email]

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George McNamara

Re: Histology Objective Lenses

In reply to this post by Neil Anthony

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Hi Neil,

I support other comments to go plan-apochromat for quality histology
images. Also a good condenser, with capability to set the condenser
numerical aperture diaphragm to mimimize glare for each objective lenses
(ideally, this would have detents to make it easy to change be touch,
without having to look all the time).

I also recommend these two scanners:

Single slide scanner: Pathscan Enabler V (consider getting the optional
polarizer, useful for histology as well as the marketed geology application

https://www.meyerinst.com/pathscan-enabler-5

"Scan area 36.5 x 24.3 mm, 14,320 x 9,522 pixels", 2.55x2.55 um pixel
size (I do not know the effective numerical aperture).

The "5" is higher quality than the earlier version used for the Original
Tiki_Goddess portrait, whose 84" 'fabric' print is now my core mascot
(digital mascot in 2nd link below)

http://home.earthlink.net/~tiki_goddess/TikiGoddess.jpg

http://confocal.jhu.edu/gallery

Flatbed scanner with full size transparency lid: Epson Perfection V850
Pro Photo Scanner ("6400 dpi, a 4.0 Dmax") ... I note 25.4 mm / 6400
pixels = ~4x4 um pixel size.

https://epson.com/For-Work/Scanners/Photo-and-Graphics/Epson-Perfection-V850-Pro-Photo-Scanner/p/B11B224201
(ignore 'out of stock' and list price)

https://www.amazon.com/Epson-Perfection-V850-Pro-scanner/dp/B00OCEJMG8
(one or many suppliers)

optional fluid mount accessory is for 'wet mount' scanning, might be
useful for 'bio scanning'

https://www.amazon.com/Epson-Fluid-Mount-Accessory-V800/dp/B00OKB52JE

You can put a whole lot of microscope slides on the Epson V850 and
acquire in "one click".

Accessorize!

Software for Epson V850:

* may come with Lasersoft Silverfast Ai (and calibration target). If so:
nicer than the Epson software.

* Hamrick VueScan Pro www.hamrick.com ... and do spend the extra couple
of dollars to get the "Pro" edition, enables modes up to "raw" 64-bit
scans (16 bit for R,G,B,IR). I like for the Epson (and pretty much any
scanner except the Pathscan Enabler, which has its own acquisition
software that works with the device).

O.D. Calibration targets (useful for V850 and other scanners, can also
be used on microscopes)

* http://www.stouffer.net/TransPage.htm

Stouffer is not the only target available ... Edmund Optics/Scientific,
ThorLabs, Reynard (but Reynard may still be selling standard only 2 mm
thick)

http://www.reynardcorp.com/anti-reflection-coatings/optical-components/fixed-neutral-density-filters/step-neutral-density-filters/33-step-linear-nd-filters-vis.html

See Jerry Sedgewick's section on OD calibration (which I had encouraged
Jerry to include)

https://books.google.com/books?id=T8l-SVgI0NMC&pg=PA48&lpg=PA48&dq=stouffer+optical+density+calibration&source=bl&ots=iIYUk_G4iF&sig=GoU1O-og7fQubrhwEV738eDVu4g&hl=en&sa=X&ved=0ahUKEwj24ZX17ubUAhVGdT4KHbsiDF4Q6AEINzAD#v=onepage&q=stouffer%20optical%20density%20calibration&f=false

old imageJ stuff

https://imagej.nih.gov/ij/docs/examples/calibration/

MetaMorph has a dropin ODscale to enable you to calculate OD in images.
Maybe someday some Fiji ImageJer will implement an equivalent in the 'J.

George
p.s. I have previously posted here recommending previous versions of
Pathscan Enabler and Epson (V750-M in the old days).

Also, while nicer to have high tungsten halogen lamp intensity (I like
11 https://www.youtube.com/watch?v=KOO5S4vxi0o ) and neutral density
filter (usually 50x50 mm glass, you could use 52 mm diameter glass
filters from amazon.com photography section), can color balance in
Photoshop if needed.

http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf

Can also acquire with scientific grade monochrome camera through "DAPI",
"GFP", and 'Red' (Texas Red probably better than Cy3) filter sets
(cubes) or emission filters (wheel). The new ORCA-Lightning 12MP sCMOS
could be fun,
http://www.hamamatsu.com/us/en/news/news/20170613000001.html

On 6/30/2017 1:10 PM, Anthony, Neil wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi all, I hope the scopes treat you well.
>
> I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.
>
> Not having much histology experience, I have to ask:
> If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?
>
> Thanks in advance for your time.
>
> Neil
>

--

George McNamara, PhD
Baltimore, MD 21231
[hidden email]
https://www.linkedin.com/in/georgemcnamara
https://works.bepress.com/gmcnamara/75 (may need to use Microsoft Edge or Firefox, rather than Google Chrome)
http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650
http://confocal.jhu.edu (as of May 22, 2017)

Neil Anthony

Re: Histology Objective Lenses

In reply to this post by Martin Wessendorf-2

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*****

Thanks Martin, good to know it's just my email local that giving the warnings.

Also, I like the idea of borrowing a set of lenses.

Thanks
Neil

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Martin Wessendorf
Sent: Friday, June 30, 2017 2:57 PM
To: [hidden email]
Subject: Re: Histology Objective Lenses

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Neil--

First, I think that the "fraudulent email" alert must be local to Emory--I've never seen it on any posting to the list, including yours!

Regarding objectives--there could be a lot of answers to this question, depending in part on your budget and on your anticipated uses. I have a turret with 6 objectives, four of which I use daily (4x, 10x, 20x, and 40x dry)--I'd have a hard time living without any of those. (I use my 2x and 60x oil lenses less often.) However, magnification doesn't tell the full story--we also need to talk about objective type (e.g.,acromats, fluorites, planapochromats, etc.), numerical aperture, transmittance at different wavelengths, etc. What you want will depend on what you will be looking at and recording.

--Could you talk to your dealer and see if they could lend you a full set for an extended period, from which you could pick the ones that you like?

Good luck!

Martin

On 6/30/2017 1:19 PM, Anthony, Neil wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> And for completeness, the below email is not fraudulent, and I think
> the auto system has a false positive. Unless of course you tell me
> your trade secrets about objective lenses that I can sell for big
> money, then things might be different ;)
>
> Neil
>
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of Anthony, Neil
> Sent: Friday, June 30, 2017 1:10 PM
> To: [hidden email]
> Subject: Histology Objective Lenses
>
> [This sender failed our fraud detection checks and may not be who they
> appear to be. Learn about spoofing at
> http://aka.ms/LearnAboutSpoofing]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi all, I hope the scopes treat you well.
>
> I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.
>
> Not having much histology experience, I have to ask:
> If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?
>
> Thanks in advance for your time.
>
> Neil
>
>
>
>
>
>
>
>
>
> ________________________________
>
> This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited.
>
> If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments).

Neil Anthony

Re: Histology Objective Lenses

In reply to this post by TSwayne

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

Thanks Theresa,
I'll think about the different sample plates too. Good to know.

Neil

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Swayne, Theresa C.
Sent: Friday, June 30, 2017 2:42 PM
To: [hidden email]
Subject: Re: Histology Objective Lenses

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Neil,

Two types of lenses we use all the time for histology are:
1) very low-mag (like 2.5x and 5x) for documenting large areas, and
2) “high-dry” (meaning high NA, no oil) like 10x/0.5, 20x/0.8 or 40x/0.8.
“Plan” (flat-field corrected) lenses give you good correction across those wide fields.

We don’t use oil on histology samples much, but if you did have the budget for an oil lens, 60x should be enough for most users and high-pixel-count cameras.

If you will have users walking in with cell culture plates or chamber slides with living cells, you might also want a long-working-distance lens… probably 20x or maybe 40x.
I would definitely get a phase ring on that lens (some vendors have phase rings in the lens, others not). A correction collar for substrate thickness will give you a sharper image on a wider variety of samples.

Hope this helps, and happy shopping!
Theresa

On Jun 30, 2017, at 2:19 PM, Anthony, Neil <[hidden email]<mailto:[hidden email]>> wrote:

Hi all, I hope the scopes treat you well.

I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.

Not having much histology experience, I have to ask:
If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?

Thanks in advance for your time.

Neil

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Confocal and Specialized Microscopy Shared Resource<http://hiccc.columbia.edu/research/sharedresources/confocal>

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Phone: 212-851-4613
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Neil Anthony

Re: Histology Objective Lenses

In reply to this post by George McNamara

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Thanks George, I appreciate the detailed answer.

I’m also intrigued by the use of flatbed scanners for whole slide imaging. Does the coverslip give troubles with interference or reflections? Also, what might you use the wet mount for?

Thanks
Neil

From: George McNamara [mailto:[hidden email]]
Sent: Friday, June 30, 2017 9:05 PM
To: Confocal Microscopy List <[hidden email]>; Anthony, Neil <[hidden email]>
Subject: Re: Histology Objective Lenses

Hi Neil,

I support other comments to go plan-apochromat for quality histology images. Also a good condenser, with capability to set the condenser numerical aperture diaphragm to mimimize glare for each objective lenses (ideally, this would have detents to make it easy to change be touch, without having to look all the time).

I also recommend these two scanners:

Single slide scanner: Pathscan Enabler V (consider getting the optional polarizer, useful for histology as well as the marketed geology application

https://www.meyerinst.com/pathscan-enabler-5

"Scan area 36.5 x 24.3 mm, 14,320 x 9,522 pixels", 2.55x2.55 um pixel size (I do not know the effective numerical aperture).

The "5" is higher quality than the earlier version used for the Original Tiki_Goddess portrait, whose 84" 'fabric' print is now my core mascot (digital mascot in 2nd link below)

http://home.earthlink.net/~tiki_goddess/TikiGoddess.jpg

http://confocal.jhu.edu/gallery

Flatbed scanner with full size transparency lid: Epson Perfection V850 Pro Photo Scanner (" 6400 dpi, a 4.0 Dmax") ... I note 25.4 mm / 6400 pixels = ~4x4 um pixel size.

https://epson.com/For-Work/Scanners/Photo-and-Graphics/Epson-Perfection-V850-Pro-Photo-Scanner/p/B11B224201 (ignore 'out of stock' and list price)

https://www.amazon.com/Epson-Perfection-V850-Pro-scanner/dp/B00OCEJMG8 (one or many suppliers)

optional fluid mount accessory is for 'wet mount' scanning, might be useful for 'bio scanning'

https://www.amazon.com/Epson-Fluid-Mount-Accessory-V800/dp/B00OKB52JE

You can put a whole lot of microscope slides on the Epson V850 and acquire in "one click".

Accessorize!

Software for Epson V850:

* may come with Lasersoft Silverfast Ai (and calibration target). If so: nicer than the Epson software.

* Hamrick VueScan Pro www.hamrick.com<http://www.hamrick.com> ... and do spend the extra couple of dollars to get the "Pro" edition, enables modes up to "raw" 64-bit scans (16 bit for R,G,B,IR). I like for the Epson (and pretty much any scanner except the Pathscan Enabler, which has its own acquisition software that works with the device).

O.D. Calibration targets (useful for V850 and other scanners, can also be used on microscopes)

* http://www.stouffer.net/TransPage.htm

Stouffer is not the only target available ... Edmund Optics/Scientific, ThorLabs, Reynard (but Reynard may still be selling standard only 2 mm thick)

http://www.reynardcorp.com/anti-reflection-coatings/optical-components/fixed-neutral-density-filters/step-neutral-density-filters/33-step-linear-nd-filters-vis.html

See Jerry Sedgewick's section on OD calibration (which I had encouraged Jerry to include)

https://books.google.com/books?id=T8l-SVgI0NMC&pg=PA48&lpg=PA48&dq=stouffer+optical+density+calibration&source=bl&ots=iIYUk_G4iF&sig=GoU1O-og7fQubrhwEV738eDVu4g&hl=en&sa=X&ved=0ahUKEwj24ZX17ubUAhVGdT4KHbsiDF4Q6AEINzAD#v=onepage&q=stouffer%20optical%20density%20calibration&f=false

old imageJ stuff

https://imagej.nih.gov/ij/docs/examples/calibration/

MetaMorph has a dropin ODscale to enable you to calculate OD in images. Maybe someday some Fiji ImageJer will implement an equivalent in the 'J.

George
p.s. I have previously posted here recommending previous versions of Pathscan Enabler and Epson (V750-M in the old days).

Also, while nicer to have high tungsten halogen lamp intensity (I like 11 https://www.youtube.com/watch?v=KOO5S4vxi0o ) and neutral density filter (usually 50x50 mm glass, you could use 52 mm diameter glass filters from amazon.com photography section), can color balance in Photoshop if needed.

http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf

Can also acquire with scientific grade monochrome camera through "DAPI", "GFP", and 'Red' (Texas Red probably better than Cy3) filter sets (cubes) or emission filters (wheel). The new ORCA-Lightning 12MP sCMOS could be fun,
http://www.hamamatsu.com/us/en/news/news/20170613000001.html

On 6/30/2017 1:10 PM, Anthony, Neil wrote:

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To join, leave or search the confocal microscopy listserv, go to:

http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy

Post images on http://www.imgur.com and include the link in your posting.

*****

Hi all, I hope the scopes treat you well.

I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.

Not having much histology experience, I have to ask:

If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?

Thanks in advance for your time.

Neil

--

George McNamara, PhD

Baltimore, MD 21231

[hidden email]<mailto:[hidden email]>

https://www.linkedin.com/in/georgemcnamara

https://works.bepress.com/gmcnamara/75 (may need to use Microsoft Edge or Firefox, rather than Google Chrome)

http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650

http://confocal.jhu.edu (as of May 22, 2017)

________________________________

This e-mail message (including any attachments) is for the sole use of
the intended recipient(s) and may contain confidential and privileged
information. If the reader of this message is not the intended
recipient, you are hereby notified that any dissemination, distribution
or copying of this message (including any attachments) is strictly
prohibited.

If you have received this message in error, please contact
the sender by reply e-mail message and destroy all copies of the
original message (including attachments).

George McNamara

Re: Histology Objective Lenses

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Neil,

I did not encounter any problems with interference fringes or reflection
with the flatbed scanner. The wet mount is an option intended for film.
Don't expect 'perfect focus' (though hard to tell with large pixel size).

George

On 7/6/2017 9:41 PM, Anthony, Neil wrote:

>
> Thanks George, I appreciate the detailed answer.
>
> I’m also intrigued by the use of flatbed scanners for whole slide
> imaging. Does the coverslip give troubles with interference or
> reflections? Also, what might you use the wet mount for?
>
> Thanks
>
> Neil
>
> *From:*George McNamara [mailto:[hidden email]]
> *Sent:* Friday, June 30, 2017 9:05 PM
> *To:* Confocal Microscopy List <[hidden email]>;
> Anthony, Neil <[hidden email]>
> *Subject:* Re: Histology Objective Lenses
>
> Hi Neil,
>
> I support other comments to go plan-apochromat for quality histology
> images. Also a good condenser, with capability to set the condenser
> numerical aperture diaphragm to mimimize glare for each objective
> lenses (ideally, this would have detents to make it easy to change be
> touch, without having to look all the time).
>
> I also recommend these two scanners:
>
> Single slide scanner: Pathscan Enabler V (consider getting the
> optional polarizer, useful for histology as well as the marketed
> geology application
>
> https://www.meyerinst.com/pathscan-enabler-5
>
> "Scan area 36.5 x 24.3 mm, 14,320 x 9,522 pixels", 2.55x2.55 um pixel
> size (I do not know the effective numerical aperture).
>
> The "5" is higher quality than the earlier version used for the
> Original Tiki_Goddess portrait, whose 84" 'fabric' print is now my
> core mascot (digital mascot in 2nd link below)
>
> http://home.earthlink.net/~tiki_goddess/TikiGoddess.jpg
> <http://home.earthlink.net/%7Etiki_goddess/TikiGoddess.jpg>
>
> http://confocal.jhu.edu/gallery
>
> Flatbed scanner with full size transparency lid: Epson Perfection V850
> Pro Photo Scanner ("6400 dpi, a 4.0 Dmax") ... I note 25.4 mm / 6400
> pixels = ~4x4 um pixel size.
>
> https://epson.com/For-Work/Scanners/Photo-and-Graphics/Epson-Perfection-V850-Pro-Photo-Scanner/p/B11B224201
> (ignore 'out of stock' and list price)
>
> https://www.amazon.com/Epson-Perfection-V850-Pro-scanner/dp/B00OCEJMG8
> (one or many suppliers)
>
> optional fluid mount accessory is for 'wet mount' scanning, might be
> useful for 'bio scanning'
>
> https://www.amazon.com/Epson-Fluid-Mount-Accessory-V800/dp/B00OKB52JE
>
> You can put a whole lot of microscope slides on the Epson V850 and
> acquire in "one click".
>
> Accessorize!
>
> Software for Epson V850:
>
> * may come with Lasersoft Silverfast Ai (and calibration target). If
> so: nicer than the Epson software.
>
> * Hamrick VueScan Pro www.hamrick.com <http://www.hamrick.com> ... and
> do spend the extra couple of dollars to get the "Pro" edition, enables
> modes up to "raw" 64-bit scans (16 bit for R,G,B,IR). I like for the
> Epson (and pretty much any scanner except the Pathscan Enabler, which
> has its own acquisition software that works with the device).
>
> O.D. Calibration targets (useful for V850 and other scanners, can also
> be used on microscopes)
>
> * http://www.stouffer.net/TransPage.htm
>
> Stouffer is not the only target available ... Edmund
> Optics/Scientific, ThorLabs, Reynard (but Reynard may still be selling
> standard only 2 mm thick)
>
> http://www.reynardcorp.com/anti-reflection-coatings/optical-components/fixed-neutral-density-filters/step-neutral-density-filters/33-step-linear-nd-filters-vis.html
>
> See Jerry Sedgewick's section on OD calibration (which I had
> encouraged Jerry to include)
>
> https://books.google.com/books?id=T8l-SVgI0NMC&pg=PA48&lpg=PA48&dq=stouffer+optical+density+calibration&source=bl&ots=iIYUk_G4iF&sig=GoU1O-og7fQubrhwEV738eDVu4g&hl=en&sa=X&ved=0ahUKEwj24ZX17ubUAhVGdT4KHbsiDF4Q6AEINzAD#v=onepage&q=stouffer%20optical%20density%20calibration&f=false
>
> old imageJ stuff
>
> https://imagej.nih.gov/ij/docs/examples/calibration/
>
> MetaMorph has a dropin ODscale to enable you to calculate OD in
> images. Maybe someday some Fiji ImageJer will implement an equivalent
> in the 'J.
>
> George
> p.s. I have previously posted here recommending previous versions of
> Pathscan Enabler and Epson (V750-M in the old days).
>
> Also, while nicer to have high tungsten halogen lamp intensity (I like
> 11 https://www.youtube.com/watch?v=KOO5S4vxi0o
> <https://www.youtube.com/watch?v=KOO5S4vxi0o> ) and neutral density
> filter (usually 50x50 mm glass, you could use 52 mm diameter glass
> filters from amazon.com photography section), can color balance in
> Photoshop if needed.
>
> http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf
> <http://home.earthlink.net/%7Egeomcnamara/McNamara2005JoH28n2pp81-88.pdf>
>
> Can also acquire with scientific grade monochrome camera through
> "DAPI", "GFP", and 'Red' (Texas Red probably better than Cy3) filter
> sets (cubes) or emission filters (wheel). The new ORCA-Lightning 12MP
> sCMOS could be fun,
> http://www.hamamatsu.com/us/en/news/news/20170613000001.html
>
>
> On 6/30/2017 1:10 PM, Anthony, Neil wrote:
>
> *****
>
> To join, leave or search the confocal microscopy listserv, go to:
>
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>
> Post images onhttp://www.imgur.com and include the link in your posting.
>
> *****
>
> Hi all, I hope the scopes treat you well.
>
> I was wondering if anybody would advise on the best objective lenses for imaging histology based slides. I'm outfitting a scope for basic widefield, check-your-sample, fluorescence, phase etc., type setup, that has a color camera for histology too.
>
> Not having much histology experience, I have to ask:
>
> If you had to pick two or three objective lenses that you couldn't do without for your histology samples, what would they be?
>
> Thanks in advance for your time.
>
> Neil
>
>
>
> --
> George McNamara, PhD
> Baltimore, MD 21231
> [hidden email] <mailto:[hidden email]>
> https://www.linkedin.com/in/georgemcnamara
> https://works.bepress.com/gmcnamara/75 (may need to use Microsoft Edge or Firefox, rather than Google Chrome)
> http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650
> http://confocal.jhu.edu (as of May 22, 2017)
>
> ------------------------------------------------------------------------
>
> This e-mail message (including any attachments) is for the sole use of
> the intended recipient(s) and may contain confidential and privileged
> information. If the reader of this message is not the intended
> recipient, you are hereby notified that any dissemination, distribution
> or copying of this message (including any attachments) is strictly
> prohibited.
>
> If you have received this message in error, please contact
> the sender by reply e-mail message and destroy all copies of the
> original message (including attachments).