Research Flow & Imaging
Cytometry Resource


 
 
Imaging Cytometry
 

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The 3-laser imaging cytometer Imagestream100 (405, 488 and 658 nm excitation lines) is located at the BL2+- laboratory (second floor, Armenise building). The acquisition and upgrade was supported by the NIH Shared Instrumentation Grant 1S10RR23459-01 and by the Immune Disease Institute funds. Current optical configuration allows the excitement and acquisition of the following dyes:


1) From the 488 nm laser:

a) Channel 3: FITC, Alexa 488, GFP, YFP, SYBR Green;

b) Channel 4: PE, Alexa 546, Alexa 555; Cy3; DsRed;

c) Channel 5: 7-AAD; Propidium iodide; Alexa 610, PE-Alexa 610;
d) Channel 6: PERCP; PERCP-Cy5.5, PE-Cy5, PE-Cy5.5, PE-Alexa 647, PE-Alexa680, Draq 5;

2) From the 658 nm laser:


a) Channel 6: Alexa 647; Alexa 680; APC; Cy5; Cy5.5;

3) From the  405 nm laser:

a) Channel 2: DAPI; Alexa 405; Alexa 430; Pacific Blue; Cascade Blue

The Imagestream 100 is the first generation of imaging flow cytometer and combine advantages of flow cytometry (the ability to interrogate statistically significant numbers of events (cells and particles)) with those of image analysis (digital imagery of single cells and the ability to quantitate morphological features; co-localisation of fluorescent probes etc.)


The FICR users in need of image cytometry analysis of adhesive cell lines and primary cells have access to laser scanning cytometers (iCyte) located at the Westwood facility of Compucyte Inc. in a collaborative agreement.

 

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The Program in Cellular and Molecular Medicine (PCMM) is an affiliate of Children's Hospital Boston and Harvard Medical School.