Methods for Live Microscopy of Drosophila Spermatocytes


The fruitfly Drosophila melanogaster offers a rich and varied source of differentiated cell types. This and its exceptional experimental tractability make it well suited for cell cycle or other developmental biology investigations. In particular, the sperm‐producing cells of the testes provide a powerful system for studying the mechanics of cell division. These meiotically dividing primary spermatocytes can be easily isolated from the testes of mutant or transgenic animals and maintained in short‐term primary cultures. The cells' large and flat geometry makes them amenable to a variety of live cell light‐microscopy‐based observation methods. Single‐plane transmitted light time‐lapse imaging and more advanced multi‐dimensional widefield or confocal fluorescence microscopy have been used to define the morphological and kinetic changes that accompany processes ranging from meiotic chromosome segregation to microtubule dynamics. The ability to document such dynamic changes underscores the power of live cell imaging in understanding cell physiology and function.

Key Concepts

  • Drosophila is an experimentally tractable source of different cell types.
  • Male meiotic primary spermatocytes provide a powerful model system for studying cell division.
  • These large flat cells are easily cultured for live cell light microscopy.
  • Transmitted light and fluorescence imaging methods can be used separately or in tandem to record different dynamic processes during cell division.

Keywords: live cell imaging; mitosis; meiosis; light microscopy; fluorescent protein

Figure 1. The primary spermatocyte culture system. (a) Testes (arrowheads) isolated from a pharate adult in phosphate buffered saline. (b) The aluminium slide and attached coverslip used as a culturing chamber. (c) Low magnification view of a culture. Primary spermatocyte cells entering meiosis (*) can be recognised by their large size and nuclear morphology. Also present in this culture are post‐meiotic cells before differentiation (arrowhead) as well as those that have matured into sperm as revealed by groups of flagella (arrow). Bar is 20 µm.
Figure 2. Selected frames from a multi‐dimensional recording of a spermatocyte with GFP‐tagged microtubules undergoing anaphase. At each time point, DIC transmitted light and widefield fluorescence images were acquired at six successive focal planes. The single centre‐most DIC optical section for each time point is shown. The corresponding fluorescence panels are the inverted superimposed projections of all six sections and reveal the distribution of microtubules throughout the cellular volume. The combination of transmitted light and fluorescence microscopy allows for the tracking of chromosomes (white arrowheads) as they segregate on individual bundles of shortening microtubules (black arrowheads). Bar is 20 µm.


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How to Cite close
Savoian, Matthew S(Jan 2015) Methods for Live Microscopy of Drosophila Spermatocytes. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0020869.pub2]