Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Laser Scanning Cytometry: Principles and Applications--An Update

doi: 10.1007/978-1-62703-056-4_11

Figure Lengend Snippet: Schematic representation of the laser scanning cytometer (LSC) (see text for explanation). It should be noted that the most recent models of LSC (iGeneration) have an inverted format with the laser illumination originating beneath the microscope slide (see ref. (15)).

Article Snippet: However, because cells are measured while suspended in a stream of liquid and subsequently discarded the analytical capability of FCM is limited for such applications as: The time-resolved events such as enzyme kinetics, drug uptake or efflux, cannot be analyzed in individual cells Morphology of the measured cell may only be assessed after sorting, which is cumbersome and not always available Subcellular localization of the fluorochrome cannot be analyzed The cell, once measured, cannot be reanalyzed with another probe(s) Analysis of solid tissue requires cell or nucleus isolation that leads to loss of information on tissue architecture Small-sized samples, such as fine needle aspirates or spinal fluid, are seldom analyzed by FCM because repeated sample centrifugations, that often are required, lead to cell loss The sample once measured is lost and cannot be stored for archival preservation The microscope-based laser scanning cytometer (LSC), designed by Kamentsky ( 1 – 3 ) and manufactured since the mid 1990s by CompuCyte Corp. (Westwood, MA), offers many of the advantages of FCM, but does not have the limitations listed above.

Techniques: Cytometry, Microscopy

Journal: Cancers

Article Title: Calcium Independent Effect of Orai1 and STIM1 in Non-Hodgkin B Cell Lymphoma Dissemination

doi: 10.3390/cancers10110402

Figure Lengend Snippet: Orai1 and STIM1 expression is altered in extra-nodal DLBCL. ( A ) Representative immunofluorescent staining of STIM1 and Orai1 in normal lymph node and extra-nodal DLBCL under-expressing STIM1 or Orai1. Images were acquired using Leica DMI8 microscope equipped with a 40× oil immersion objective. TMA including samples from normal lymph node (n = 26), nodal (n = 43) and extra-nodal (n = 44) DLBCL was co-immunostained using Orai1 or STIM1 antibodies revealed by donkey anti-rabbit Alexa-488 (in green) and mouse anti-human CD20 and anti-human CD19 revealed by goat anti-mouse Alexa 532 (in red). Nuclei were stained with DAPI (in blue). Scale bar = 50 µm. ( B ) Quantification of STIM1 or Orai1 alteration in DLBCL samples. After acquisition of spot fluorescence on Icys laser scanning cytometer, a segmentation analysis base on phantoms was done to determine the percentage of CD20/CD19 positive cells expressing protein of interest for each spot. The stacked column charts illustrate the intensity grade for STIM1 or Orai1 expression in surgical specimens. * p < 0.05 by Chi-square test.

Article Snippet: After acquisition on Icys laser scanning cytometer (Thorlabs, Maison Lafitte, France), a segmentation analysis base on phantoms was done to determine the percentage of CD20/CD19 positive cells expressing protein of interest for each spot as described previously [ ].

Techniques: Expressing, Staining, Microscopy, Fluorescence, Cytometry