In This Issue
Western New York Flow Users Group
FEATURED NEW PRODUCTS
what it is
how it works
|Tali™ Image-Based Cytometer||1 instrument||T10796|
|Tali™ Apoptosis Kit—Annexin V Alexa Fluor® 488 and Propidium Iodide||1 kit||A10788|
what they are
how they work
|Product ||Quantity ||Cat. No.|
|Click-iT® EdU Alexa Fluor® 488 Flow Cytometry Assay Kit ||50 assays||C10425|
|Click-iT® EdU Alexa Fluor® 647 Flow Cytometry Assay Kit ||50 assays||C10424|
|Click-iT® EdU Pacific Blue™ Flow Cytometry Assay Kit ||50 assays||C10418|
New Protein Disulfide Isomerase (PDI) Endoplasmic Reticulum Marker — ABfinity™ Recombinant Rabbit Monoclonal Antibodies
what they are
how they work
|Are my cells alive? Proliferating? How did my compound affect cell viability? |
Is my compound cytotoxic? Researchers across many areas of cellular biology ask these questions when planning and performing experiments. Cell health (viability, proliferation, cytotoxicity, and cell death) is one of the more complex areas of cell biology, and answers can be provided by a wide variety of assays for various aspects of cell structure and function. Common cell health indicators measure reduction potential/metabolic status, membrane integrity, induction of proteases, loss of mitochondrial membrane potential, dsDNA breaks, nucleic acid content, and more.
No single assay can give a definitive answer on the health of a cell, and it is therefore common to use more than one approach when assessing cellular health at any given time point. Cell health assays inform important decisions in both academic and drug discovery research, and unambiguous answers are essential. To help answer such questions with more confidence, we have developed multiparametric cell health assays that offer:
- Learn More About HCS Cell Health and Cytotoxicity Assays
|Imaging of mitotoxicity and cytotoxicity of valinomycin in HeLa cells using the HCS Mitochondrial Health Kit. HeLa cells were treated in a dose-response experiment with valinomycin between 2 nM and 120 μM final concentrations or with an equal volume of DMSO (control), incubated for 24 hr at 37°C/5% CO2, and assayed using the HCS Mitochondrial Health Kit. |
The Thermo Scientific Cellomics® ArrayScan® VTI platform was used to obtain images of fixed cells at 20x. In the absence of valinomycin, the nucleus (detected with Hoechst 33342) and mitochondrial reticulum (detected with MitoHealth stain) were clearly visible, yet no signal due to permeability of plasma membranes (Image-iT® DEAD Green™ viability stain) was observed. At valinomycin concentrations in the nanomolar range, the mitochondrial reticulum was not observed as a defined structure, indicating valinomycin-induced loss of mitochondrial membrane potential. Loss of plasma membrane integrity did not occur at this level of valinomycin exposure, hence there was no fluorescence staining by the Image-iT® DEAD Green™ viability stain. At higher valinomycin concentrations (120 μM), however, plasma membrane integrity was compromised and resulted in intense nuclear fluorescence from the Image-iT® DEAD Green™ viability stain, indicating more overt cytotoxicity and cell death.
Narita M, Young AR, Arakawa S et al. (2011) Science 332:966–970.
In a recent publication, Narita et al. propose a model by which cells can achieve both protein synthesis and autophagic degradation simultaneously. Using Premo™ Autophagy Sensor LC3B-RFP to investigate autophagy, CellLight® Golgi-GFP to localize and study the function of the Golgi complex, and Click-iT® Homopropargylglycine (HPG) to detect nascent protein synthesis, the researchers observed that, in cells undergoing senescence, autophagosomes fuse with mTORC1-containing lysosomes to form a specialized compartment (TASCC) for the execution of the secretory program. Because this compartment enforces a close proximity between the autophagy process and mTORC1, the amino acids and metabolites produced during autophagy activate mTORC1 to boost protein synthesis. These results demonstrate that cells can, under certain conditions, couple mass breakdown (catabolism via autophagy) to mass production (anabolism via protein synthesis) through the formation of a novel compartment, the TASCC.
- View the Bibliography Reference
The Molecular Probes® Handbook
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