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Flow Cytometry01:23

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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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IR Frequency Region: X–H Stretching01:24

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In IR spectroscopy, signals produced by the X−H bonds (such as C−H, O−H, or N−H) can be observed in the frequency range of  2700–4000 cm–1. The C−H stretching vibration forms sharp bands in the region 2850–3000 cm–1. The presence of the O−H stretching vibration leads to the forming of an absorption band in the frequency range 3650–3200 cm−1. At the same time, N−H stretching can be confirmed by absorption bands in...
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Double bonds in alkenes and carbonyl compounds exhibit stretching frequencies in the diagnostic region of the IR spectrum. In addition, alkenes exhibit vinylic C–H stretching and C–H out-of-plane bending absorptions that are useful for identifying substitution patterns.
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Imaging Biological Samples with Optical Microscopy01:18

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy ATOM
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High-throughput imaging flow cytometry by optofluidic time-stretch microscopy.

Cheng Lei1, Hirofumi Kobayashi2, Yi Wu2,3

  • 1Department of Chemistry, The University of Tokyo, Tokyo, Japan. leicheng@chem.s.u-tokyo.ac.jp.

Nature Protocols
|July 7, 2018
PubMed
Summary
This summary is machine-generated.

Optofluidic time-stretch microscopy enables high-throughput imaging flow cytometry for analyzing cellular heterogeneity. This method allows for rapid, large-scale single-cell analysis across diverse cell types, advancing biological and medical research.

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Area of Science:

  • Biotechnology
  • Microscopy
  • Cell Biology

Background:

  • Understanding cellular heterogeneity is crucial for biological and medical advancements.
  • High-throughput methods are needed to analyze large cell populations.
  • Existing techniques may lack the speed or resolution for comprehensive single-cell analysis.

Purpose of the Study:

  • To provide a detailed protocol for optofluidic time-stretch microscopy.
  • To enable high-throughput, single-cell imaging flow cytometry.
  • To facilitate large-scale analysis of cellular morphological and molecular variations.

Main Methods:

  • Construction of an optical time-stretch microscope and a microfluidic device.
  • High-throughput single-cell image acquisition at >10,000 cells/s with sub-micrometer resolution.
  • Image construction, enhancement, and analysis using computational tools like compressive sensing and machine learning.

Main Results:

  • Demonstration of a protocol for optofluidic time-stretch microscopy.
  • Achieved high-throughput imaging flow cytometry for diverse cell types.
  • Enabled large-scale single-cell analysis of cellular heterogeneity.

Conclusions:

  • Optofluidic time-stretch microscopy is a powerful tool for high-throughput single-cell analysis.
  • This protocol facilitates research in biology, medicine, pharmaceuticals, and green energy.
  • The method addresses the challenge of analyzing 'big data' in cellular research.