Methods of Studying Cells

5 Key excerpts on "Methods of Studying Cells"

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  eBook - PDF

  Molecular Cytology V1

  The Cell Cycle

  • Jean Brachet(Author)
  • 2012(Publication Date)
  • Academic Press

    (Publisher)

  CHAPTER 2 TECHNIQUES USED IN CELL BIOLOGY: A BRIEF SUMMARY The study of cell physiology calls for a combination of many different meth-ods. It would be fruitless to present them here in detail since new techniques are being continuously introduced, while classical methods are still undergoing im-provements and modifications. All possible aspects of the methodology used in cell biology can be found in the publication, Methods in Cell Biology. Be-sides the description of techniques for cell culture, cell separation, cell fusion, nuclear transplantation, dissection of cells in nucleate and anucleate halves, and others, this series contains much useful information on older methods that remain of fundamental importance: optical methods, electron microscopy, cytochemical techniques, and the isolation of cell constituents by differential centrifugation of homogenates. Their merits will be discussed only briefly here. I. OPTICAL METHODS The classical technique of light microscopy remains the basis of any cell study, whether the observations are made on living cells (sometimes vitally stained with appropriate dyes) or on sections (or smears) of fixed and stained tissues. Video microscopy greatly enhances the definition of the structures seen under the light microscope. There are several useful complements to traditional light microscopy (see Barer, 1966, for a review). Phase contrast microscopy provides improved con-trast for the study of living, transparent cells; it is particularly useful when beating of cilia or flagella, cell movement, or cell division should be followed by microcinematography. Interference microscopy also shows details (especially with Nomarski's interference-contrast optics that cannot be seen under a light microscope and has the additional advantage of enabling one to measure the dry mass of the cells (Davies et al. y 1953). Polarization microscopy is useful for the study of birefringent objects such as mitotic or meiotic spindles.

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  Cells and Tissues

  An Introduction to Histology and Cell Biology

  • Rogers(Author)
  • 2012(Publication Date)
  • Academic Press

    (Publisher)

  2 The techniques available In a very obvious sense, the techniques that exist for studying cells and tissues determine our knowledge. The very existence of cells was unknown until lenses were developed which permitted them to be seen. Until the development of electron microscopy, the ultrastruc-ture of the cell was a mystery. But the influence of techniques on our knowledge is more far-reaching than that. Histology grew into a science in the second half of the nineteenth century, a time which saw steady improvement in the performance of the light microscope, and also the appearance of a chemical industry which, particularly in Germany, was often directed to producing new textile dyes. Sections prepared then provided the basis for the descriptions and classifications of cells which we still use today. Each new combination of cell shape, size, orientation, position and staining characteristics became a named cell type. The relationships between various cell types were continually and often bitterly debated, because standard histological techniques did not permit evidence to be collected on transformations between one cell type and another. So cells that look alike, such as lymphocytes, have a common name, though we now know that this cell type includes cells with at least two very different life histories. Cells that look different have different names, though we now know that, for instance, the B-lymphocyte and the plasma cell are different stages in the life of the same cell. All through histology run references to basophilic (loving basic dyes), acidophilic (loving acidic dyes), chromophobe (not staining with common dyes) and metachromatic (staining, but having a colour that is different from that of the original dye). Cells can be described and classified in many different ways which are valid and useful. The classification we use grew out of the techniques available when the detailed study of tissues and organs began.

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  Histochemical and Cytochemical Methods of Visualization

  • Jean-Marie Exbrayat(Author)
  • 2013(Publication Date)
  • CRC Press

    (Publisher)

  1.1.6 Q UANTITATIVE M ETHODS Since the 1980s, new methods for observation and analysis appeared, using a computer to quantify the images. Flow cytometry is used to study cells one by one, using the light absorbed or emitted by the cells, the natural fluorescence, or that of a specific fluorescent dye. Flow cytometry also permits one to pick out the cells according to the chosen parameters. In automatic quantitative analysis, the observations are done by a camera, then analyzed by means of a computer. This method allows colorimetric studies, spatial reconstruction, and measur-ing without subjectivity. 1.2 GENERAL PRINCIPLES OF HISTOLOGY AND HISTOCHEMISTRY To perform a histological study, tissues and cells must be submitted to a series of operations allow-ing preservation and visualization of organic components with a light microscope. For electron microscopy study, tissues also must be submitted to a series of similar operations (see Chapter 8). Histology gives a general view of tissue structure, often using empirical methods. With histo-chemistry methods, the nature of chemical components of tissues and cells can be precise, accord-ing to a specific staining. In this case, staining reactions are known with sufficient precision and one may modify parameters according to the component researched.

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  eBook - ePub

  The Cell

  • Britannica Educational Publishing, Kara Rogers(Authors)
  • 2010(Publication Date)
  • Britannica Educational Publishing

    (Publisher)

  CHAPTER 6The Study of Cells

  T he study of cells as fundamental units of living things forms the basis of the field known as cell biology. The earliest phase of cell study began with English scientist Robert Hooke’s microscopic investigations of cork and his introduction of the term cell in 1665. In the 19th century two Germans, the botanist Matthias Schleiden and the biologist Theodor Schwann, were among the first to clearly state that cells are the fundamental particles of both plants and animals. This pronouncement—the cell theory—was amply confirmed and elaborated by a series of discoveries and interpretations.

  In 1892 the German embryologist and anatomist Oscar Hertwig suggested that organism processes are reflections of cellular processes. His work established cytology (now generally referred to as cell biology) as a separate branch of biology. Research into the activities of chromosomes led to the founding of cytogenetics, in 1902–04, when the American geneticist Walter Sutton and the German zoologist Theodor Boveri demonstrated the connection between cell division and heredity.

  Modern cell biologists have adapted many methods of physics and chemistry to investigate cellular events. Improvements in techniques for growing cells in the laboratory have revolutionized science and medicine. For example, scientists now can build “bioartificial” tissues for transplantation into patients and are able to investigate individual steps in the process of cell differentiation. These developments have important implications in medicine, specifically for the regeneration of tissue in persons affected by certain diseases.

  THE HISTORY OF CELL THEORY

  Although the microscopists of the 17th century had made detailed descriptions of plant and animal structure and Hooke had coined the term cell

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  Practical Cell Analysis

  • Dimitri Pappas, Dimitri Pappas(Authors)
  • 2010(Publication Date)
  • Wiley

    (Publisher)

  This chapter has outlined many of the techniques available for cell microscopy, as well as many practical solutions to culturing cells on the microscope and obtaining the best possible image. Many of the techniques discussed in this chapter, such CONCLUSION 121 as staining/fixing protocols, can be used in flow cytometry or other techniques. When combined with the protocols and probes listed in Chapter 9, the optical microscope can be used to elucidate most cell processes with relative ease and with high information content. REFERENCES 1. Byassee, T.A., Chan, W.C.W., and Nie, S. (2000) Probing single molecules in single living cells. Analytical Chemistry , 72 , 5606–5611. 2. Kohl, T., Haustein, E., and Schwille, P. (2004) Determining protease activity in vivo by fluorescence cross-correlation analysis. Biophysical Journal , 89 , 2770–2782. 3. Zhang, J., Fu, Y., Liang, D. et al. Fluorescent avidin-bound silver particle: a strategy for single target molecule detection on a cell membrane. Analytical Chemistry , 81 , 883–889. 4. Nguyen, Q.-T., Callamaras, N., Hsieh, C., and Parker, I. (2001) Construction of a two-photon microscope for video-rate Ca 2 þ imaging. Cell Calcium , 30 , 383–393. 5. Callamaras, N. and Parker, I. (1999) Construction of a confocal microscope for real-time X-Y and X-Z imaging. Cell Calcium , 26 , 271–279. 6. Liu, K., Dang, D., Harrington, T. et al. (2008) Cell culture chip with low-shear mass transport. Langmuir , 24 , 5955–5960. 7. Liu, K., Tian, Y., Pitchimani, R. et al. (2009) Characterization of PDMS-modified glass from cast-and-peel fabrication. Talanta , 79 , 333–338. 8. Howell, J.L. and Truant, R. (2002) Live-cell nucleocytoplasmic protein shuttle asay utilizing laser confocal microscopy and FRAP. Biotechniques , 32 , 80–87. 9. Hwang, E.Y., Pappas, D., Jeevarajan, A.S., and Anderson, M.M. (2004) Evaluation of the paratrend multi-parameter sensor for potential utilization in long-duration auto-mated cell culture monitoring.

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