New leukocyte quantification method - human & large animal blood

Dr. Ina Laura Pieper & colleagues (Cytometry Part A (2016). DOI: 10.1002/cyto.a.22874) from Swansea University and Calon Cardio describe the combined use of CyTRAK Orange and DRAQ7 to robustly quantify leukocytes in whole blood samples.  

CyTRAK Orange (Em max 610 nm) was used to gate the nucleated cells while DRAQ7 (Em max 694 nm) was used to quantify dead / damaged cells, alongside counting beads.

The methodology was specifically developed to monitor in human, bovine and ovine samples for in vitro testing of heart pumps (left ventricular assist devices, LVAD) where cell damage is a key concern, though it should be generally applicable.  

Such testing is not possible by haematology analyser and requires the capabilities of a flow cytometer.

The combined use of CyTRAK Orange and DRAQ7 permits the additional use of violet-excited chromophores, FITC and APC for more detailed phenotypic analysis.

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Chagas' Disease: searching for therapeutics with HCS

Two recent publications describe the use of DRAQ5 to aid the continuing search for new drugs to defeat infection by T. Cruzi.  

DRAQ5 powerfully allowed identification of individual host cells by nuclear staining, secondary segmentation of their cytoplasm and identification and enumeration of the intracellular parasites by labelling kinetoplast DNA in a single fluorescence channel!  

The methodology is compatible with high content screening (HCS) automation and automated image analysis for the identification of positive hits that inhibit parasitic infection.  

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References: 

Alonso-Padilla, Julio, et al. 

"Automated high-content assay for compounds selectively toxic to Trypanosoma cruzi in a myoblastic cell line." 

PLoS Neglected Tropical Diseases 9.1 (2015): e0003493.

& 

Silva, Fredson T., et al. 

"Design, synthesis and antitrypanosomal activity of some nitrofurazone 1, 2, 4-triazolic bioisosteric analogues." 

European Journal of Medicinal Chemistry (2016). Jun 1. 

Accepted, in press.

Seeing the whole heart - developmental biology

Stunning images of the whole developing vertebrate (Xenopus) heart should enable new understanding of the exquisite and tightly controlled process that are required to achieve the fully functioning heart chambers from what starts as a simple tube of muscle.

The lab of Tim Mohan -The Francis Crick Institute - used CyGEL Sustain to immobilise the delicate whole tissue for fluorescence microscopy.

Based on the core patented technology of CyGEL as a unique thermo-reversible mountant for live cells and tissues, CyGEL Sustain can be formulated with your culture medium of choice to achieve optimal conditions.  CyGEL gels upon warming above room temperature, gently immobilizing live objects for high-performance imaging. Simple cooling of CyGEL then allows non-destructive recovery of your precious living sample(s) for downstream onward development or orthogonal analysis.

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Reference:

Smith, Stuart J., et al. "The cardiac-restricted protein ADP-ribosylhydrolase-like 1 is essential for heart chamber outgrowth and acts on muscle actin filament assembly." Developmental biology (2016).

doi:10.1016/j.ydbio.2016.05.006

Find out more about CyGEL Sustain at:
http://www.biostatus.com/CyGEL-Sustain/