2016 schedule

Lab 1 1 DAPI slide per pair

Lab 2

  • same DAPI slide as Lab 1

  • 1 H&E slide from histology

Lab 3 Numerical aperture . same DAPI slide

Lab 4 Resolution.

  • 1 slide newly stained phalloidin actin. Must be bright

  • 1 slide fluorescent beads(PS-Speck Microscope Point Source Kit P7220 from Molecular Probes)

Lab 5 Fluorescence. 1 new, bright 3-color stained slide (DAPI, Phalloidin-actin, FITC microtubules)

Lab 6 Dry lab

Lab 7 Labeling Cells 9/28/16

  • 3 fixed LL coverslips

  • 1 3t3 fixed coverslip

  • 1 live LL coverslip for students to fix

  • paraglut

  • forceps, mounting medium, pasteur pipettes

Lab 8 - Organelles 10/3/16

  • Golgi - anti-58K, mouse, formaldehyde fixation

  • Kinetochore - anti Hec1 - MeOH fixation 1:200 mouse

  • ER anti-calnexin, paraformaldehyde fixation 1:200

  • centrosome - anti gamma-tubulin, paraglut, mouse 1:100

  • nuclear envelope anti LAP2 - MeOH fixation 1:100 mouse

  • lysosomes

  • alpha actinin - mouse 1:200 formaldehyde fixation

Lab 8 Practical (and pipet practice)- 10/5

Lab 9 - Imaging Live Cells 10/11 TUESDAY Live cells on mattek

unknown GFP tagged proteins

Thaw GFP lines Wednesday
Split Thursday (~1/4)
Plate Sunday on Mateks for Tuesday

  • Thaw
    • GFP tubulin
    • actin thaw
    • EB1 - thaw
  • Nucleofect: EGFP-H2B
    • split LL parentals on Sunday, HEAVY
    • nucleofect & plate Monday by noon

Lab 10 10/12

  • Single cells to whole organisms

    • order protists

    • induce GFP E. coli

  • CRISPR: Transform guide RNA plasmid into bacteria

Lab 11.1 Motility I 10/17

  • thaw B16, 3t3 Thursday 10/13/16 (late in the day)
  • plate 9 MatTeks each Fri 10/14/16:
    • 3t3
    • actin GFP LL
    • B16

Lab 11.2 Motility II 10/19

Lab 11.3 Motility III 10/24

Motility presentations 10/26

CRISPR: prepare Cas9/guide RNA plasmid DNA 10/31

CRISPR: prepare repair DNA cassette (PCR, clean up) 11/2

CRISPR: Add Cas9 plasmid + repair DNA to cells 11/7

Lab 13.1 - Cell Signaling 11/9

Lab 13.2 - Cell Signaling 2 11/14

11/16 Wed = Friday schedule

Discussion of independent projects 11/28

Projects 11/30 - 12/14

Final exam period: Final project presentation

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