Mitosis Through the Microscope: Advances in Seeing Inside Live Dividing Cells Conly L. Rieder and Alexey Khodjakov

Figure S1
Microtubules at the edge of a newt lung cell visualized, after fixation and staining, by video-enhanced DIC (A, A’) and indirect immunofluorescence (B,B’) LM. The boxed region shown in A and B is seen at higher magnification in A’ and B’. Note that although individual microtubules are visible in the more cluttered DIC image, they are much easier to follow in the fluorescence image.

Linked websites of labs that work on mitosis: Many contain pictures, movies, etc.
http://www.bio.unc.edu/faculty/salmon/lab/mitosis/mitosislabs.html
Linked websites of labs that work on cytokinesis:
http://www.bio.unc.edu/faculty/salmon/lab/mafia/mafialabs.html

To view these movies, download a QuickTime viewer.

• Movie S1
  Time-lapse video light microscopy of a newt lung epithelial cell, undergoing the metaphase to anaphase transition, as seen by the Oldenbourg 360°r; polarized system. In these images the spindle appears white. [courtesy of Rudolf Oldenbourg (Marine Biology Laboratory, Woods Hole, MA USA) and Phong Tran, Dept. of Biology, Univ. of N. Carolina, Chapel Hill, N.C., USA) .

• Movie S2
  Time-lapse polarization video light microscopy of several primary crane fly spermatocytes entering and completing meiosis I. As the spindle forms in these cells the kinetochore fibers appear as prominent white bundles that connect each chromosome to the spindle poles. The stem body also is clearly visible as it forms during cytokinesis, [courtesy of Rudolf Oldenbourg (Marine Biology Laboratory, Woods Hole, MA USA) and James R. LaFountain (Department of Biology, State Univ. of New York, Buffalo, Buffalo, N.Y. USA)].

• Movie S3
  Time-lapse DIC video light microscopy of spindle formation and chromosome congression in a PtK₁ cell. After nuclear envelope breakdown (6 sec into the movie) several chromosomes mono-orient to the top and bottom poles of the forming spindle. One of these exhibits rapid poleward motion (7 seconds). Over the next 28 seconds all of these mono-oriented chromosomes become bioriented and move to the spindle equator, to establish the metaphase plate. In the movie 1 second is equivalent to approximately 1 minute in real time. (courtesy of Alexey Khodjakov, Lab of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York USA).

• Movie S4
  Time-lapse video DIC light microscopy of a chromosome as it attaches to the prometaphase spindle during in a newt lung cell. In this example one chromosome (at 7 o’clock) suddenly exhibits rapid motion ~ 1 second into the movie towards the closest centrosome (at 11 o’clock) . (courtesy of Conly L. Rieder, Lab of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York USA)

• Movie S5
  Time-lapse video-enhanced DIC light microscopy of a growing astral microtubule (arrowhead from 1 sec on) contacting one of the kinetochores on an unattached chromosome. As a result of this contact the chromosome becomes attached to one of the spindle poles, which it subsequently moves rapidly towards. (courtesy of Conly L. Rieder, Lab of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York USA).

• Movie S6
  Time-lapse video DIC light microscopy showing how chromosomes behave during meiosis I in a wildtype Drosophila spermatocyte. In this example the persistent nucleolus, defined as the refractive spherical organelle in the nucleus at the start of the movie, is expelled towards the left-hand pole of the forming spindle ~ 3 seconds into the movie. By this time all of the chromosomes have achieved a metaphase alignment, and anaphase starts 8 seconds into the movie. Time in hrs:min:sec, is seen in upper left hand corner of the movie (courtesy of Conly L. Rieder and Matthew Savoian, Lab of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York USA)

• Movie S7
  Time-lapse video DIC light microscopy of how chromosomes behave during anaphase I in a Drosophila primary spermatocyte mutant for the kinetochore protein, zw- 10. Note that the rate of poleward motion is severely attenuated relative to that seen in wildtype (Movie S6) spermatocytes, and also that the chromosomes decondense well prior to reaching the polar regions. Time in hrs:min:sec, is seen in upper left hand corner of the movie (courtesy of Conly L. Rieder and Matthew Savoian, Lab of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York USA).

• Movie S8
  Recovery of fluorescence after photobleaching the -tubulin-GFP associated with one of the centrosomes in a PtK₁ cell blocked in mitosis by nocodazole treatment. The left hand panel shows the centrosomes as seen by epi-fluorescence, while the right panel shows the cell and chromosomes as seen by DIC. The upper centrosome (3 o’clock) is photobleached at 9:57 AM, and slowly regains its fluorescence intensity over the next several hours. Time, in hrs:min:sec, is in lower right hand corner of each frame. (courtesy of Alexey Khodjakov, Lab of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York USA).

• Movie S9
  Speckled fluorescence microscopy imaging of a spindle formed in vitro in Xenopus oocyte extracts. This film reveals that tubulin subunits within spindle microtubules are constantly moving (fluxing) poleward. (courtesy of Tarun Kapoor, Rockefeller University, New York, N.Y., USA).

• Movie S10
  Time-lapse multi-mode light microscopy of mitosis in a cancer (HT1080) cell transfected with histone H2B-GFP. In this example the cell is followed with both epifluorescence (left) and phase-contrast (right) light microscopy. The former reveals the behavior of the chromosomes during spindle formation, anaphase and telophase, while the latter depicts the dynamic properties of the rest of the cell, especially during cytokinesis. Each frame represents a maximum intensity projections of 17 optical sections, and 5950 fluorescence images were used to construct the film (courtesy of Dr. Olga Kisurina-Evgenieva, Lab of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York USA).