RE: [Histonet] Flourescent labels

From:"Glenn Smith"


I know one of our customers use our TISSUEscope instrument to scan
immunofluorescence and immunohistochemistry (RGB) slides and had imaged
fluorescence ffpe tissues. When I asked her about your question, her
comments to me regarding the TISSUEscope, were that formalin fixed paraffin
embedded is no different from the frozen sections. 

Our systems employ the narrow bandwidth filters that Tim Morken speaks of
below and are laser-based. The specific system cited above uses blue
(488nm), green (532nm) and red (635nm) laser sources (with relevant filters)
to digitally capture the entire sample in either brightfield (rgb) or
fluoresence contrast modes.  We have other customer labs that have different
laser combinations depending on the fluorophores of interest and have
supplied a variety of sources between 400 - 800 nm (violet, blue, green,
yellow, red, ruby) matched up with the appropriate filter combinations.


Glenn Smith, P.Eng.
ph: 519.886.9013 x38
mobile: 519.498.0614

Biomedical Photometrics Inc./GeneFocus
A12-550 Parkside Dr.
Waterloo, ON N2L 5V4
Widefield Laser Scanning Instruments and Software for Fluorescence and
Brightfield Imaging

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Message: 19
Date: Fri, 12 Aug 2005 16:31:52 -0400
From: "Morken, Tim - Labvision" 
Subject: RE: [Histonet] Flourescent labels
To: 'Patsy Ruegg' , 'Histonet'
Content-Type: text/plain


The average routine fluorescent system uses "longpass" emission filters that
let almost all the light from blue to red through. Therefore you will see
many colors of  autofluoresce. Since the autofluorescence is usually a
different color than your target fluorchrome, the way you can avoid
autofluoresence is to find filters that block the autofluorescent
wavelength. It takes more effort to coordinate the different wavelengths
associated with the fluorochrome excitation and emission vs the
autofluorescent excitation and emission, but by using narrow-bandwidth
filters (20-40 nm bandwidth) it is usually possible. Both Omega and Chroma
have these filters.

Tim Morken
Lab Vision - Neomarkers

-----Original Message-----
[] On Behalf Of Patsy Ruegg
Sent: Friday, August 12, 2005 1:18 PM
To: 'Histonet'
Subject: [Histonet] Flourescent labels

I was under the general impression that most flourescent labels were done on
frozen tissue because of problems with auto flourescence associated with
aldehyde fixation and paraffin processing.  Has the technology developed so
that these new flourescent labels such as the Alexa dyes can now be used on
ffpe tissues? Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 216
Aurora, CO 80010
wk email
web site  

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