Triplet cell division of a pancreatic tumor cell (43 hours, full field, 50% zoom)

This is

http://works.bepress.com/gmcnamara/60

and is a full field of view, and ~20 hours longer than the movie I posted yesterday at

http://works.bepress.com/gmcnamara/59/

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I previously posted a triplet cell division movie (from years ago) at

http://works.bepress.com/gmcnamara/21/

on a triplet cell division of "U251t glioma cell division to triplets, acquired in 2005 by Prof. Laurence Cooper, Dr. Cludia Kowolik and George McNamara, at City of Hope National Medical Center, Duarte, CA, using CoH's Video Timelapse Light Microscopy Facility (VTLM). 3 hour video, one frame every two minutes." (that video was probably acquired with a 10x objective lens).

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Prof. Cooper moved to Houston in 2006, I moved to Miami in 2007, and Cooper recruited me to Houston in 2013. Now i Sept 2014 I am doing timelapse imaging of pancreatic tumor cells -- in preparation for adding antigen specific cytolytic T-lymphocytes (CTLs, aka 'killer T-cells') to observe and quantify "serial killing". During imaging of this experiment our colleague, Dr. Ana Korngold, pointed out this triplet cell division (mid-field of view, about 50% of the way through this video). My thanks to Dr. Shan Zong, Hiroki Torikai and Amir Alpert, in our lab for this collaboration - especially Shan for providing these cells.

Due to our experiment program, I cannot provide any details of the pancreatic tumor cells used here. The entire triplet division took about 90 minutes, the normal (2 daughter) cell divisions may be faster.

Time interval: 2 minutes.

Playback: 10 frames per second.

Time duration: 2590 min (1295 frames * 2 min interval), equals 43.166 hours.

Magnification 20x.

Pixel size is 1000 nm (1 um), because I made a 50% zoomed stack in MetaMorph from the original 500 nm (0.5 um).

Field of view 1024x1024 pixels due to zooming 50% from 2048x2048 pixels in the original data, This also means the X and Y axes are 1024x1024 um.

Camera: Hamamatsu FLASH4.0 sCMOS (2,048x2048 pixel camera, zoomed 50% for this movie).

Microscope: Leica DMI6000.

Objective lens and mode: HCX PL Fluotar 20x/0.40NA and phase contrast

(I use a Leica HC Plan Apo 20x/0.75 when doing fluorescence imaging, this dataset is only transmitted light so I used the phase contrast objective lens and imaging mode).

Live cell incubator: PECON.

Imaging dish: Mattek 35 mm diameter, 20 mm diameter imaging area, #1.5 coverglass.

Culture media: DMEM with Glutamax (3 mL).

Video format: AVI file using Intel IYUV format.

As noted above, this is the "entire" 43.166 hour dataset (2048x2048 pixels, still 2 min interval).

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This movie (bepress 60) has (at least) 3 unconventional cell divisions. The triplet division at bottom center about 300 frames (~1/4 through), and later triplet or other bizarre divisions on the right side and near top.

George McNamara, PhD

Single Cells Analyst / Sr Research Scientist

L.J.N. Cooper lab

MD Anderson Cancer Center

Houston, TX

p.s. Dr. Ana Korngold also noticed something often seen with human cells in tissue culture: the nuclei and nucleoli sometimes look like human faces. I have posted about this previously, see

http://works.bepress.com/gmcnamara/25/

for a Halloween "trick or treat" package, where you can self-identify which cell looks like you, your friends, your ex-friends if you point out certain resemblances, your colleagues, etc (great to do during committee meetings, if you run out of faces, try self-identifying and committee-member identifying by psychogeometrics using this score card

http://www.willamette.edu/dept/careers/pdf/Psychogeometrics%20Willamette%202010.pdf

(stay on page 1 until you have identified everyone!)