Method/Kit:
Microscopy - Fluorescence

Measurement Type Imaging
Description Imaging with some unspecified number of fluorescent marker and/or stains.

Target/Analytes with this Method/Kit
Target/Analyte Short Name Description
Actin Filaments Actin Filaments Actin Filaments - typically shown in an image. Example of stain to view actin filaments is FITC-Phalloidin.
Antibody diffusion Antibody diffusion Fluorescence antibody diffusion will be monitoring at several time points.
Calcium Transients - 1Hz Ca2+_1Hz Ca2+ transient amplitudes were calculated as the maximum relative change in fluorescence signal, ΔF/F = (Peak − Trough)/(Trough − Background).
Calcium Transients - 2Hz Ca2+_2Hz Ca2+ transient amplitudes were calculated as the maximum relative change in fluorescence signal, ΔF/F = (Peak − Trough)/(Trough − Background).
Calcium Transients - Tetanus Ca2+_Tetanus Ca2+ transient amplitudes were calculated as the maximum relative change in fluorescence signal, ΔF/F = (Peak − Trough)/(Trough − Background).
Calcium Transients - Twitch Ca2+_Twitch Ca2+ transient amplitudes were calculated as the maximum relative change in fluorescence signal, ΔF/F = (Peak − Trough)/(Trough − Background).
Cell Growth - Population Doubling Time PD Cells were imaged at 4, 24, 48, 72, and 96 hours on a Nikon Eclipse TE2000-U microscope at 10x magnification using the NIS Elements software and analyzed using ImageJ. The log of percent growth over time was plotted and using the slope of the line of best fit, population doubling time (td) was calculated by the following formula: td = ln(2)/(slope*2.303)
E-Selectin E-Selectin Quantification of mean fluorescence intensity (MFI) of the images in following the recovery for 8 days at 0.5 ml/min in TEBVs for E-Selectin
ICAM-1 ICAM-1 Quantification of mean fluorescence intensity (MFI) of the images in following the recovery for 8 days at 0.5 ml/min in TEBVs for ICAM-1
Image IMG Image for later analysis of intensity, morphology, distribution, location, etc.
Monocyte Accumulation Rate Monocyte/cm^2 To measure monocyte accumulation and migration, monocytes were labeled with cell tracker red-CMTPX (Life Tech) before addition to the perfusion loop. At the end of monocyte perfusion, the TEBV lumen was washed with PBS gently to remove unattached cells. Then the TEBVs were fixed with 4% Paraformaldehyde (PFA) solution for one hour. The TEBV is opened en face and compressed between two coverslips. The accumulation of monocytes is analyzed based on fluorescence images and the migration of monocytes is analyzed from confocal images at different depths into the TEBV from the endothelial surface. For
Myoblast Purity Purity Purity - For each image, myoblast purity was calculated as the total number of vimentin+ cells subtracted from the total number of cells myoblast (desmin+, vimentin+) and fibroblast (vimentin+) cell
Trop2 Trop-2 Trop-2 is an epithelial marker for prostate cancer
VCAM-I VCAM-I Quantification of mean fluorescence intensity (MFI) of the images in following the recovery for 8 days at 0.5 ml/min in TEBVs for VCAM-1
6-CF Uptake (Image based) 6-CF Uptake (image) Uptake of 6-CF (OAT Substrate), quantified by fluorescent image analysis
Aquaporin 1 AQP1 Aquaporin 1
Cell Morphology Cell Morphology Imaging, typically with transmitted light, to see the cell morphology.
EAM-1 Uptake (image based) EAM-1 Uptake (image) Uptake of EAM-1 (OCT substrate), quantified by fluorescent image analysis
FITC-Dextran FITCdex Direct read of FITC-Dextran in media effluent typically to assess permeability.
Image IMG Image for later analysis of intensity, morphology, distribution, location, etc.
Live Cells LIVE Number or percent of live cells
MRP2/4 Efflux MRP2/4 Measurement of transport activity via MRP2 and 4
Nuclei Nuclei Nuclei as observed by image staining.
P-gp P-gp Efflux transporter
SGLT2 SGLT2 Glucose transporter
Tumor Cells Cells Used when imaging and looking at tumor cells, or counting tumor cells, or any such assay where Tumor Cells are the assay target.
ZO-1 ZO-1 Recommended name: Tight junction protein ZO-1 Alternative name(s): Tight junction protein 1 Zona occludens protein 1 Zonula occludens protein 1 TJP1, TJP2, and TJP3 are closely related scaffolding proteins that link tight junction (TJ) transmembrane proteins such as claudins, junctional adhesion molecules, and occludin to the actin cytoskeleton (PubMed:7798316, PubMed:9792688). [https://www.uniprot.org/uniprot/Q07157]

Studies Utilizing this Method/Kit
Study Start Date Study Types Description
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