By Natasha S. Barteneva, Ivan A. Vorobjev
This certain quantity for the 1st time explores strategies and protocols related to quantitative imaging circulation cytometry (IFC), which has revolutionized our skill to research cells, mobile clusters, and populations in a extraordinary style. starting with an creation to expertise, the e-book maintains with sections addressing protocols for experiences at the telephone nucleus, nucleic acids, and FISH strategies utilizing an IFC software, immune reaction research and drug screening, IFC protocols for apoptosis and cellphone loss of life research, in addition to morphological research and the id of infrequent cells. Written for the hugely winning Methods in Molecular Biology sequence, chapters contain introductions to their respective issues, lists of the required fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and pointers on troubleshooting and heading off identified pitfalls.
Authoritative and sensible, Imaging circulate Cytometry: tools and Protocols might be a severe resource for all laboratories trying to enforce IFC of their study studies.
Read Online or Download Imaging Flow Cytometry: Methods and Protocols PDF
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Additional info for Imaging Flow Cytometry: Methods and Protocols
Sample text
9): 1. The microchannel is fabricated by curing PDMS on a silicon mold. The silicon mold is prepared by soft lithography. 2. First, a layer of photoresist is coated on a silicon wafer using a spin coater. 3. The photoresist is soft baked at 65 and 95 °C for 3 and 6 min on a hot plate, respectively. 4. The silicon wafer and photoresist are then cooled under ambient temperature before being exposed to ultraviolet (UV) light. 5. A maskless soft lithography machine is used to transfer a pattern defined by a computer-aided design (CAD) software onto the photoresist.
C, d) The corresponding line profiles (yellow dotted lines) of the single-angle ATOM images in a, b, respectively. Each line scan of the image is captured within ~4 ns. , a + b), an image with absorption contrast can be revealed. , a − b). (g) White-light DIC image of the same MIHA cell for comparison. e. e. on-axis fiber coupling), in the context of high-speed flow imaging (8 m/s) of the MIHA cells in a PDMS microfluidic channel. , BF time-stretch image (right). The bottom inset is the corresponding line profiles (yellow dotted lines) of ATOM image and BF timestretch image shown.
Incubator (Thermo Scientific, Heratherm, IGS60). 4. Expanded plasma cleaner (Harrick Plasma, PDC-002). 5. Cover slips. 6. Polymethyl methacrylate (PMMA). 7. Maskless soft lithography machine Patterning, LLC SF-100 XCEL). (Intelligent Micro 8. Photoresist (MicroChem Corp. SU-8 2025). 9. Spin coater (Apex Instruments Co. spin NXG-P1). 10. Hot plate (IKA, C-MAG HP 7). 11. SU-8 developer. 12. Isopropyl alcohol. 13. PDMS precursor (SYLGARD 184 Silicone Elastomer): 18 g Sylgard 184 Silicone elastomer oligomer and 2 g cross-linker were mixed together by an agitator.