Flow Cytometry is in some way not a classical hard science. There are sometimes several ways to get the answer for the question you asked. However there are some rules that are good to follow. It is in some way like cooking, there a lot of recipes out there for spaghetti sauce. Whereas a simple mistake might spoil your meal.
We therefore offer again a flow cytometry day this year. For the advanced users to refresh their knowledge on current practices in flow cytometry and for the novice users to get a comprehensive introduction to flow cytometry. And we will have a presentation of a new technique for the simultaneous analysis of multiple cytokines in the afternoon. And we are again happy to have Thomas Bauer form Ayoxxa here for presentation.
We start in the morning with the general introduction to flow cytometry and expand the Agenda to Multicolor, Data Aquisition, Apoptosis and Bead Based Assays. You can join us any time you like and there will be ample time for discussion during the sessions and the breaks. So here is the program as a .pdf printout and the agenda in details.
Date: Tuesday 25. August, 2015
Location: Uniklinikum, Biomedizinisches Zentrum, Kleiner Hörsaal
And please leave us a note (fccf@uni-bonn.de), if you want to participate, so that we can take care of coffee and food.
| 10:00–11:30 | Flow Cytometry: Principles, Instrument Set up, Optics, Filter, Staining (IC/EC) |
| 11:30-11:45 | Coffee Break |
| 11:45–12:30 | Multicolor Flow Cytometry; Fluorescent Dyes, their properties, selection, sensitivities |
| 12:30–13:30 | Lunch Break |
| 13:30–14:30 | How to optimize Flow Cytometry data acquisition and experimental design to receive optimal results |
| 14:30–14:45 | Coffee Break |
| 14:45-15:30 | Apoptosis detection and functional assays: Viability, Vitality, Proliferation…. |
| 15:30–16:30 | A novel way of Multiple Cytonkien Detection: 2-dimensional readout in a 3-dimensional assay |
The ongoing trend in increasing fluorescence options for Flow Cytometry as well as Imaging Instruments demands for the usage of a broad range of fluorescent dyes. This covers typically the Ultraviolet- (350nm), Violet- (405nm), Blue- (488nm), Yellow-Green (561nm) and Red- (630nm) – laser lines as excitation source. According to the individual experiment requirements the selection of fluorescent dyes towards the target needs to be carefully qualified especially when a multicolour analysis is desired (8 and more color experiment). The different key dyes will be discussed towards their individual characteristic like brightness, molecular weight and application.
Flow Cytometry experiments require careful consideration of many aspects; on the instrument side, the choice of dyes, biological context and staining procedures to receive optimal sensitivity. We will have a look into ways to optimize the experiment to receive best resolution by: exclusion of false positive signals, doublet discrimination, reduction of background as well as dead cell exclusion including the application on fixed cell, all with the intention to deliver best possible results. Furthermore we will discuss bead compensation, experimental standardization and fluorescence channel optimization.
Apoptosis is a highly regulated and evolutionary conserved pathway of cell death that plays a critical role in development and maintenance of tissue homeostasis. Apoptosis progression in mammals can be broken down into a number of key steps: induction, activation, and execution. We will have a look at a cellular example and detect the different stages of apoptosis on a time depended manner by demonstrating the different assays associated beginning with mitochondrial changes and to end with controlled cellular disintegration.
Multiple analyte detection is an invaluable tool for the comprehensive study of biological systems, as they are comprised of networks of cytokines, chemokines, growth factors and related proteins. “The parallel detection of multiple cytokines in various sample types becomes more and more essential to understand complex biological responses and processes. AYOXXA Biosystems has developed a new, proprietary detection platform based on chip technology, providing exceptional robustness, simplicity in use and readout using 3µl sample volume only.
See you and for any questions do not hesitate to contact us
Thomas Bauer (Ayoxxa, thomas.bauer@ayoxxa.com)
and your Core Facility Staff (fccf@uni.-bonn.de)
