Flow cytometry detection of surface and intracellular antigens in pancreas from a single mouse embryo

Summary: We here report a flow-cytometry-based protocol to measure single-cell protein expression in small samples. The protocol is optimized for simultaneous detection of fluorescent proteins and intracellular and surface antigens in the embryonic pancreas from the mouse. Owing to low cell numbers, current protocols for flow cytometric analysis of embryonic tissues rely on tissue pooling. Our protocol enables analysis of one pancreas per sample, thereby facilitating detection of biological variation and minimizing the number of experimental animals needed.For complete details on the use and execution of this protocol, please refer to Nyeng et al (2019)

Cyt‐Geist: Current and Future Challenges in Cytometry: Reports of the CYTO 2019 Conference Workshops

The need for cytometry instrumentation, reagents, training and scientific collaborations in the nations of Africa remains high despite strong efforts by both the African and foreign biomedical and cytometry research communities. Dr. Tesfa and Dr. Blanco therefore organized the first Cytometry in Africa Workshop at CYTO2019. This workshop had several goals. The first goal was to present the results of a pre-workshop survey aimed at assessing flow cytometry resources, personnel, experience and training in Africa. The results of this survey demonstrated important strengths in the African cytometry community, but also pinpointed areas where instrument access, reagent availability and training could be improved. The second goal was to present several collaborative scientific projects in Africa with participation by ISAC members. Third, both existing and proposed strategies for improving collaborative efforts and research support were presented, including cytometer donations, research collaborations and training programs. Finally, an open roundtable discussion was held with workshop attendees, many with experience in working in Africa. A diverse array of investigators from government, academia and industry attended and contributed to the workshop. A key outcome of the workshop was the establishment an African Working group in collaboration with the ISAC Instruments 4 Science Task Force, the ISAC Live Education Task Force, and the ISAC Education Committee. The workshop also marked the establishment of I4S, with the goal of advancing flow cytometry in the international research communit

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion