Imaging Flow Cytometry: Methods and Protocols by Natasha S. Barteneva, Ivan A. Vorobjev

By Natasha S. Barteneva, Ivan A. Vorobjev

This specified quantity for the 1st time explores options and protocols regarding quantitative imaging stream cytometry (IFC), which has revolutionized our skill to research cells, mobile clusters, and populations in a amazing type. starting with an advent to expertise, the ebook keeps with sections addressing protocols for experiences at the telephone nucleus, nucleic acids, and FISH strategies utilizing an IFC device, immune reaction research and drug screening, IFC protocols for apoptosis and mobile dying research, in addition to morphological research and the id of infrequent cells.  Written for the hugely winning Methods in Molecular Biology sequence, chapters comprise introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, simply reproducible laboratory protocols, and tips about troubleshooting and heading off identified pitfalls.

Authoritative and useful, Imaging movement Cytometry: equipment and Protocols can be a severe resource for all laboratories trying to enforce IFC of their examine studies.

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C) Further zoomed-in view of a single line-scan. Arrows indicate the positions of the two channel side-walls. 7). (e, f) Dual-contrast ATOM images: differential phase-gradient contrast and absorption contrast images formed by arithmetic subtraction and summation of dual-angle ATOM images 3. Goldstein’s algorithm is used to unwrap the phase in the wrapped phase [24]. 4. The slanted phase profile is then removed by subtracting the average phase profile in order to obtain the final quantitative phase image (Fig.

11a). 2. The high-pass-filtered interferogram is then Hilberttransformed to obtain the wrapped phase from a complex analytical signal (Fig. 11b) [23]. Ultrafast Microfluidic Cellular Imaging 39 Fig. 10 (a) Raw data of ATOM. Arrows indicates the detected individual cells. (b) Zoomed-in view of the timestretch pulses. Each pulse represents a single line in a 2D time-stretch image. Each line is digitally stacked to form the 2D image, as shown in (e). (c) Further zoomed-in view of a single line-scan.

Ultrafast Microfluidic Cellular Imaging 37 6. Afterwards, the photoresist undergoes a developing step by immersing the photoresist-coated silicon wafer in SU-8 developer for 5 min. The silicon wafer is rinsed with isopropyl alcohol followed by a drying step of either nitrogen blow or air blow over the wafer. 7. The PDMS precursor is mixed with the corresponding curing agent with a ratio of 10:1 before pouring onto the silicon mold. 8. A PMMA block is designed and placed on the silicon wafer to form a thin PDMS layer in the imaging section of the channel.

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