question: camera for macroscope

[Rietdorf, Jens]

Dear colleagues,

we are looking for suitable illumination & camera/s for our Leica Z6
MacroFluo. It is used for both fluorescence and absorption contrast
samples.

Has anybody out there experience with LED illumination in combination
with a monochrome camera to generate 'true color' images?

Your advice is welcome, commercial replies off list, please.

Thank you, jens

---
dr. jens rietdorf
head microscopy facility
novartis research foundation
friedrich-miescher-institute, wro1066.2.16
maulbeerstr.66, 4058 basel, switzerland
txt: rietdorf at fmi dot ch
fon: +41.61.69.75172
fax: [phone]

[Peter Gabriel Pitrone]

Hello Jens,

You would need a large amount of pixels to digitally resolve your  
images with such low magnification, and a large chip to collect light  
from fluorescence. That is a difficult thing to find. One interesting  
color imaging technique is made by a company called Foveon: http://
www.foveon.com/ They make a 14 megapixel 3 layer CMOS chip (called  
Foveon X3) that is 21x14 mm, that might be useful to you in this  
regard. Here is a link to the PDF: http://www.foveon.com/files/ 
F13_image_sensor_Product_Flier_RevD.pdf

I have no affiliation with the company, and have not even tried it  
out, but am intrigued by the technology.

You could also get a normal monochrome CCD Camera with an electronic  
LCD trigger, like Diagnostic Instruments SPOT RT Slider from Visitron  
GmbH or QImaging Regina with a trigger from Chromaphor.

Same disclaimer as above.

Good luck on your search!

Pete

On Oct 28, 2009, at 2:23 PM, Rietdorf, Jens wrote:

Dear colleagues,

we are looking for suitable illumination & camera/s for our Leica Z6
MacroFluo. It is used for both fluorescence and absorption contrast
samples.

Has anybody out there experience with LED illumination in combination
with a monochrome camera to generate 'true color' images?

Your advice is welcome, commercial replies off list, please.

Thank you, jens

---
dr. jens rietdorf
head microscopy facility
novartis research foundation
friedrich-miescher-institute, wro1066.2.16
maulbeerstr.66, 4058 basel, switzerland
txt: rietdorf at fmi dot ch
fon: +41.61.69.75172
fax: [phone]

[Martin Wessendorf-2]

Dear Jens--

Rietdorf, Jens wrote:

> we are looking for suitable illumination & camera/s for our Leica Z6
> MacroFluo. It is used for both fluorescence and absorption contrast
> samples.
>
> Has anybody out there experience with LED illumination in combination
> with a monochrome camera to generate 'true color' images?

Clever idea!  Sounds as if it'd work nicely for transmitted illumination
and tinctorial staining.  However, unless I'm misunderstanding you, I
don't think it would work as well for fluorescence, since the emission
wavelengths observed would be a function of the molecule rather than the
excitation wavelength.

Good luck--

Martin Wessendorf
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[Patrick Van Oostveldt]

We got good results in microscopy with leds and RGB filters obtained  
from Agfa Gevaert.
These filters were used in earlier days to print and adapt colorphotos  
and put in lightpath of the magnifier to compensate color.
They are named gelatine color filters and are availbale in different  
densities.
Probably you can also use the color head from a photographic enlarger  
if available.
You should test the exposure for different colors on a white paper and  
rescale the RGB images to obtain a white field. ImageJ allows this on  
the spot.

Bye

Patrick Van Oostveldt

  pictuesQuoting "Rietdorf, Jens" <[hidden email]>:

> Dear colleagues,
>
> we are looking for suitable illumination & camera/s for our Leica Z6
> MacroFluo. It is used for both fluorescence and absorption contrast
> samples.
>
> Has anybody out there experience with LED illumination in combination
> with a monochrome camera to generate 'true color' images?
>
> Your advice is welcome, commercial replies off list, please.
>
> Thank you, jens
>
> ---
> dr. jens rietdorf
> head microscopy facility
> novartis research foundation
> friedrich-miescher-institute, wro1066.2.16
> maulbeerstr.66, 4058 basel, switzerland
> txt: rietdorf at fmi dot ch
> fon: +41.61.69.75172
> fax: [phone]
>

--
Dep. Moleculaire Biotechnologie
Coupure links 653
B 9000 GENT

tel [phone]
fax [phone]

[Rietdorf, Jens]

Dear Martin and others,

sorry, I see that I should be more clear. Indeed the question is if
someone has experience in replacing a color camera by a combination of
multi-LED illumination and a monochrome camera.

We would prefer a monochrome camera, because this could then also be
used for efficient fluorescence imaging and we would not have to switch
between a color and a monochrome camera.

For fluorescence we are using regular epi-illumination, for LED pseudo
color we would use transmission illumination.

Thanks, jens

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Martin Wessendorf
Sent: Wednesday, October 28, 2009 3:46 PM
To: [hidden email]
Subject: Re: question: camera for macroscope

Dear Jens--

Rietdorf, Jens wrote:

> we are looking for suitable illumination & camera/s for our Leica Z6
> MacroFluo. It is used for both fluorescence and absorption contrast
> samples.
>
> Has anybody out there experience with LED illumination in combination
> with a monochrome camera to generate 'true color' images?

Clever idea!  Sounds as if it'd work nicely for transmitted illumination

and tinctorial staining.  However, unless I'm misunderstanding you, I
don't think it would work as well for fluorescence, since the emission
wavelengths observed would be a function of the molecule rather than the

excitation wavelength.

Good luck--

Martin Wessendorf
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[Peter Humphreys]

We have an old optronics magnafire camera that has 3 colour filters
(R,G,B) and blank for fluorescence.
there are similar still on the market.
With a white light LED the colour balance should only have to be set
once.

Peter

Centre for Stem Cell Research
Cambridge
UK

> Dear Martin and others,
>
> sorry, I see that I should be more clear. Indeed the question is if
> someone has experience in replacing a color camera by a combination of
> multi-LED illumination and a monochrome camera.
>
> We would prefer a monochrome camera, because this could then also be
> used for efficient fluorescence imaging and we would not have to switch
> between a color and a monochrome camera.
>
> For fluorescence we are using regular epi-illumination, for LED pseudo
> color we would use transmission illumination.
>
> Thanks, jens
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Martin Wessendorf
> Sent: Wednesday, October 28, 2009 3:46 PM
> To: [hidden email]
> Subject: Re: question: camera for macroscope
>
> Dear Jens--
>
> Rietdorf, Jens wrote:
>
> > we are looking for suitable illumination & camera/s for our Leica Z6
> > MacroFluo. It is used for both fluorescence and absorption contrast
> > samples.
> >
> > Has anybody out there experience with LED illumination in combination
> > with a monochrome camera to generate 'true color' images?
>
> Clever idea!  Sounds as if it'd work nicely for transmitted illumination
>
> and tinctorial staining.  However, unless I'm misunderstanding you, I
> don't think it would work as well for fluorescence, since the emission
> wavelengths observed would be a function of the molecule rather than the
>
> excitation wavelength.
>
> Good luck--
>
> Martin Wessendorf

[Jerry (Gerald) Sedgewick]

I haven't exactly done this application, but it appears that there are
two ways to accomplish your end.  Either you can use white light LEDs
and then rotate red, green and blue filters in front of your camera, or
you can choose combinations of LED lights to accomplish approximately
the same end.  I would advise the use of the former.

R,G, B filters in front of cameras are fairly broad bandpass filters
with some overlap (B=375nm to 525nm, G=475nm-600nm and R=600nm-675nm).  
The wavelengths that emit from colored light LEDs, on the other hand,
are narrow gaussian profiles.  While one could conceivably purchase a
series of LEDs to fill out a wavelength range (e.g. for the blue
wavelength range the following LEDs could be purchased: 385, 405, 455,
470 and 505nm), this would be expensive and still would not provide the
broad illumination of R, G, B filter set with a white light source.

Also, the white light LEDs do not emit at 500nm, but this is probably a
moot point for capturing generic color images.

Jerry

Rietdorf, Jens wrote:

> Dear Martin and others,
>
> sorry, I see that I should be more clear. Indeed the question is if
> someone has experience in replacing a color camera by a combination of
> multi-LED illumination and a monochrome camera.
>
> We would prefer a monochrome camera, because this could then also be
> used for efficient fluorescence imaging and we would not have to switch
> between a color and a monochrome camera.
>
> For fluorescence we are using regular epi-illumination, for LED pseudo
> color we would use transmission illumination.
>
> Thanks, jens
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Martin Wessendorf
> Sent: Wednesday, October 28, 2009 3:46 PM
> To: [hidden email]
> Subject: Re: question: camera for macroscope
>
> Dear Jens--
>
> Rietdorf, Jens wrote:
>
>  
>> we are looking for suitable illumination & camera/s for our Leica Z6
>> MacroFluo. It is used for both fluorescence and absorption contrast
>> samples.
>>
>> Has anybody out there experience with LED illumination in combination
>> with a monochrome camera to generate 'true color' images?
>>    
>
> Clever idea!  Sounds as if it'd work nicely for transmitted illumination
>
> and tinctorial staining.  However, unless I'm misunderstanding you, I
> don't think it would work as well for fluorescence, since the emission
> wavelengths observed would be a function of the molecule rather than the
>
> excitation wavelength.
>
> Good luck--
>
> Martin Wessendorf
>  

--
Jerry (Gerald) Sedgewick
Core Facility Director, Biomedical Image Processing Lab (BIPL)
University of Minnesota, Department of Neuroscience
1-205 Hasselmo Hall
312 Church St. S.E.
Minneapolis, MN  55455
612-624-6607
[hidden email]
http://www.bipl.umn.edu
Author: "Scientific Imaging with Photoshop: Methods, Measurement and Output."

Rawlight.com (dba "Sedgewick Initiatives")
965 Cromwell Avenue
Saint Paul, MN  55114
651-788-2261
[hidden email]
http://www.quickphotoshop.com
http://www.rawlight.com
http://www.jerrysedgewick.com

[Carol Heckman]

There is a camera by QImaging that sounds ideal for this.  It is monochrome but uses a filter wheel with liquid crystal filters to get the bands in R, G, and B.  We purchased one and will know soon how it looks.  It looked good at the demo.
Carol Heckman
Center for Microscopy & Microanalysis
Bowling Green State University
________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Jerry (Gerald) Sedgewick [[hidden email]]
Sent: Wednesday, October 28, 2009 2:15 PM
To: [hidden email]
Subject: Re: question: camera for macroscope

I haven't exactly done this application, but it appears that there are
two ways to accomplish your end.  Either you can use white light LEDs
and then rotate red, green and blue filters in front of your camera, or
you can choose combinations of LED lights to accomplish approximately
the same end.  I would advise the use of the former.

R,G, B filters in front of cameras are fairly broad bandpass filters
with some overlap (B=375nm to 525nm, G=475nm-600nm and R=600nm-675nm).
The wavelengths that emit from colored light LEDs, on the other hand,
are narrow gaussian profiles.  While one could conceivably purchase a
series of LEDs to fill out a wavelength range (e.g. for the blue
wavelength range the following LEDs could be purchased: 385, 405, 455,
470 and 505nm), this would be expensive and still would not provide the
broad illumination of R, G, B filter set with a white light source.

Also, the white light LEDs do not emit at 500nm, but this is probably a
moot point for capturing generic color images.

Jerry

Rietdorf, Jens wrote:

> Dear Martin and others,
>
> sorry, I see that I should be more clear. Indeed the question is if
> someone has experience in replacing a color camera by a combination of
> multi-LED illumination and a monochrome camera.
>
> We would prefer a monochrome camera, because this could then also be
> used for efficient fluorescence imaging and we would not have to switch
> between a color and a monochrome camera.
>
> For fluorescence we are using regular epi-illumination, for LED pseudo
> color we would use transmission illumination.
>
> Thanks, jens
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Martin Wessendorf
> Sent: Wednesday, October 28, 2009 3:46 PM
> To: [hidden email]
> Subject: Re: question: camera for macroscope
>
> Dear Jens--
>
> Rietdorf, Jens wrote:
>
>
>> we are looking for suitable illumination & camera/s for our Leica Z6
>> MacroFluo. It is used for both fluorescence and absorption contrast
>> samples.
>>
>> Has anybody out there experience with LED illumination in combination
>> with a monochrome camera to generate 'true color' images?
>>
>
> Clever idea!  Sounds as if it'd work nicely for transmitted illumination
>
> and tinctorial staining.  However, unless I'm misunderstanding you, I
> don't think it would work as well for fluorescence, since the emission
> wavelengths observed would be a function of the molecule rather than the
>
> excitation wavelength.
>
> Good luck--
>
> Martin Wessendorf
>

--
Jerry (Gerald) Sedgewick
Core Facility Director, Biomedical Image Processing Lab (BIPL)
University of Minnesota, Department of Neuroscience
1-205 Hasselmo Hall
312 Church St. S.E.
Minneapolis, MN  55455
612-624-6607
[hidden email]
http://www.bipl.umn.edu
Author: "Scientific Imaging with Photoshop: Methods, Measurement and Output."

Rawlight.com (dba "Sedgewick Initiatives")
965 Cromwell Avenue
Saint Paul, MN  55114
651-788-2261
[hidden email]
http://www.quickphotoshop.com
http://www.rawlight.com
http://www.jerrysedgewick.com

[Jerry (Gerald) Sedgewick]

Does anyone have experience with viewing necrotic tissues using DIC?  
I'm wondering how tough it'll be to differentiate between live tissues
and necrotic areas of tissue.  I'm hoping to use a 0.70NA APO lens for
DIC as well as some fluorescent images on the specimens without having
to swap out the lenses, but I have to be able to pick out and avoid the
necrotic areas of tissue.

Thanks,

Will Yutzy

--
Will Yutzy
[hidden email] <mailto:[hidden email]>, [hidden email]
<mailto:[hidden email]>
>
>  


--
Jerry (Gerald) Sedgewick
Core Facility Director, Biomedical Image Processing Lab (BIPL)
University of Minnesota, Department of Neuroscience
1-205 Hasselmo Hall
312 Church St. S.E.
Minneapolis, MN  55455
612-624-6607
[hidden email]
http://www.bipl.umn.edu
Author: "Scientific Imaging with Photoshop: Methods, Measurement and Output."

Rawlight.com (dba "Sedgewick Initiatives")
965 Cromwell Avenue
Saint Paul, MN  55114
651-788-2261
[hidden email]
http://www.quickphotoshop.com
http://www.rawlight.com
http://www.jerrysedgewick.com