http://confocal-microscopy-list.275.s1.nabble.com/Re-Confocal-calibration-performance-assesment-vendor-response-tp7587243p7587248.html
most specimens, so shading is not quite the same (advantage: confocal!).
Brightness: LED epi-illuminator power can be decreased. ND filters can
be put in excitation path for marc lamps and metal halide lamps. sCMOS
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>
> Chroma slides may be too bright for wide-field (though, to tell the truth, I don't see any need to monitor the performance of lamp-based microscopes), I don't know about Molecular Probes' slides. But one can sometimes use Chroma slides in an "unintended" way, something like a red slide with blue illumination, and then the brightness becomes more reasonable.
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of George McNamara
> Sent: Saturday, September 2, 2017 1:55 PM
> To:
[hidden email]
> Subject: Fluor-Ref slides ... pick up a box or case from Chroma at a conference ... Re: Confocal calibration & performance assesment **vendor response**
>
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> Dear listserv,
>
> Fluor-Ref slides ... pick up a box or case from Chroma at a conference.
>
> As for Thermo: how about giving away THIN fluorescent plastic specimens .. can include your logo (and no, it would not fall under the category of pens or coffee cups that you can no longer give out to U.S. academic medical center staff).
>
> Back to Chroma slides ... if not going to 'Neuo' or 'Cell Bio', and cannot wait for M&M 8/2018 in Baltimore, as an alternative you can use a brightly colored clipboard(s). Come ot think of it, Thermo could give out branded clipboards as big reference slides.
>
> enjoy,
>
> George
>
>
> On 9/1/2017 1:26 PM, Kilgore, Jason A. wrote:
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>>
>>
>> ** Vendor response **
>>
>> Hi, Javier,
>>
>> For a uniform fluorescence standard, I recommend the Fluor-Ref slides.
>> These are sold or re-sold by various places and are fairly
>> inexpensive. These are plastic slides that are autofluorescent in
>> four different wavelengths and are incredibly uniform and photostable.
>> The drawbacks are that they are extremely bright, so much so that you
>> often either have to throw in neutral density filters or image an
>> off-wavelength slide. Also, they are thicker than a standard slide.
>> Here's the webpage from Ted Pella:
>>
https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.te>> dpella.com%2Fhisto_html%2Ffluor.htm.aspx&data=01%7C01%7Cmmodel%40KENT.
>> EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26
>> 134%7C1&sdata=BcsiKMcDnxqoK1jIMqN2mZHgKzYkaN%2BzWhCmiq3iyCA%3D&reserve
>> d=0 or resold as a set of four from Fisher Scientific:
>>
https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.f>> ishersci.com%2Fshop%2Fproducts%2FNC0158348%2Fnc0158348%23%3Fkeyword%3D
>> FLUOR-REF%2BSET%2BOF%2B4%2BSLIDES&data=01%7C01%7Cmmodel%40KENT.EDU%7C3
>> 6cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1
>> &sdata=xv9a4R6u7RpajF1wKTSSuvTx5cB71pgNea6XQrt8RKg%3D&reserved=0
>>
>> There are other published methods for uniformity slides (such as a simple solution of fluorescein under a coverslip, or putting dye in agar) or labeling coated slides, but nothing comes close to the Fluor-Ref slides for photostability, storage stability, and uniformity of intensity.
>>
>> There is a good and fairly inexpensive option for some of the other needs you mention: FocalCheck microspheres slides. There are different slides, but probably Slide #1 is best for your needs, catalog F36909. The top row has options where you align rings of different colors to check alignment, or rings and cores of the microspheres. There are also TetraSpeck beads of three different sizes (4, 1, and 0.5 um) that are broadly fluorescent, which you can use for PSF as well as alignment. The bottom row has 6um beads of five different relative intensities, for doing intensity calibrations.
>> Webpage from Thermo Fisher Scientific:
>>
https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.t>> hermofisher.com%2Forder%2Fcatalog%2Fproduct%2FF36909&data=01%7C01%7Cmm
>> odel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018
>> f73e7dd15f26134%7C1&sdata=IbU37oBjbMd9zZE1RKzb4Lp8g6TWlZjE3FJAc6lxtJ0%
>> 3D&reserved=0
>> Manual:
>>
https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fasset>> s.thermofisher.com%2FTFS-Assets%2FLSG%2Fmanuals%2Fmp36909.pdf&data=01%
>> 7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a
>> 1ec44d018f73e7dd15f26134%7C1&sdata=rBOAKsVfx3HvTl0Y2kif8s9lVGzwfgfHYZc
>> %2BYsl1mY0%3D&reserved=0
>>
>> We sell fluorescent beads of smaller sizes, too, if you need them (such as 20 nm) of different wavelengths, for PSF measures.
>>
>> I hope this helps, but please feel free to contact me offline if you wish more info (such as a PowerPoint I have showing images from that last slide).
>>
>> Jason
>>
>>
>> Jason A. Kilgore
>> Technical Application Scientist
>> Molecular Probes / EVOS Tech Support
>> Thermo Fisher Scientific
>>
>> 1-800-955-6288 then option 4, then option 3, then option 2.
>> Or dial direct at +1 541 335 0353
>>
[hidden email]
>>
>> This communication is intended solely for the individual/entity to whom it is addressed. It may contain confidential or legally privileged information. Any unauthorized disclosure or copying is prohibited and may be unlawful. If you have received this communication in error, please notify the sender immediately and delete it from your system.
>>
>>
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:
[hidden email]] On Behalf Of Fco. Javier
>> Díez Guerra
>> Sent: Wednesday, August 30, 2017 5:10 AM
>> To:
[hidden email]
>> Subject: Confocal calibration & performance assesment
>>
>> *****
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>> &reserved=0 Post images on
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>>
>> Dear confocalists,
>>
>> I would like to know about the preferred methods used by facility
>> managers to regularly asses the performance of their confocal (and
>> wide-field) microscopes. In other words, which are the most used and reliable methods to check different features of confocal and wide-field microscope peformance?.
>>
>> Particularly, we'd like to check:
>>
>> - illumination uniformity across FOV, now we use a slide coated with fluorescent secondary antibody.
>>
>> - XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now we use 1um beads that can be excited with all laser lines.
>>
>> - XYZ Resolution, we use smaller beads (0,17um) to build and analyze
>> PSFs (for each wavelength)
>>
>> - Laser power and stability: for power, we use a power meter from Newport placed at the objective exit, for stability, we capture long time-series in reflexion mode with an empty preparation.
>>
>> - We do not know how to check detector (PMT, GAsP, Hybrid, etc) sensitivity and SNR.
>>
>> Thanks for your input.
>>
>>
>> --
>> Fco. Javier Diez-Guerra, PhD
>>
>> Profesor Titular UAM
>> Servicio de Microscopía Confocal
>> Centro de Biologia Molecular Severo Ochoa C/ Nicolás Cabrera, 1 Campus
>> de Cantoblanco
>> 28049 Madrid
>> SPAIN
>>
>> Tel +34 91 196 4612
>> e-mail:
[hidden email]