Bulk plan
| dates | |
|---|---|
| 08/18-29 | HeLa cells and IF 10+ slides’ (DNA, actin, microtubules) ready for first class |
| 09/2-11 | use these HeLa slides |
| 09/16 | Grow MEFs for IF 09/23 |
| Sep 23 | IF all set up, help pre-process a slide per student (fix and permeabilize) – thinking 3X 12 slides |
| Sep 25 (Th) or Sep 29 (M) | make imaging dishes to use 09/30 (T) live cell |
| 09/30 and 10/2 | need 10 imaging and 10 passaging dishes; (possible grow bacteria for plasmid prep) |
| 10/2 | POSSIBLE? Plasmid prep? |
| ?(MAYBE)? 10/3 | transfect cells to image 10/7 |
| 10/7-9 | Kills off cells and get new next week, Drew gone this week |
| Week of 10/13 | we will hand you new cells in prep for CURE IF #1 10/16 (Th) and 10/21 (T), for each we will want 2X slide per student…maybe more |
| Depending on how CURE goes, the rest is unplanned | |
Course Schedule – subject to change during semester
| Week | class | date | questions & themes | to do |
|---|---|---|---|---|
| PRE | 08/18-29 | HeLa IF for 10+ slides’ DNA, actin, microtubules | ||
| 1 | 1 | 09/02 | Learn your microscope | use HeLa IF, classes 1-6 |
| 2 | 09/04 | Transmitted light and alignment | ||
| 2 | 3 | 09/09 | Fluorescence | |
| 4 | 09/11 | Fluorescence photobleaching | ||
| 3 | 5 | 09/16 | Cameras, SNR, and calibration | Micrometer ***PREPARE, MEFs growing for IF next week, keep growing |
| 6 | 09/18 | Numerical Aperture and resolution | ||
| 4 | 7 | 09/23 | Immunofluorescence procedure Primaries from Drew’s lab: – H3K9me2/3 Mouse CST 5327s – H3K27me3 Rabbit CST 9733s Secondaries: – Goat anti-mouse -red – Goat anti-rabbit green | Pre-process IF slides: Fix and permeabilize |
| 8 | 09/25 | Immunofluorescence imaging | Students need 10 flasks to split into 2 flasks and an imaging dish. recent MEF slides, FOCUS learning CURE | |
| 5 | 9 | 09/30 | Live cell imaging and cell culture | 10 imaging dishes (made the previous class) 10 flasks for passaging |
| 10 | 10/02 | Plasmid prep. and cell culture | 10 student passaging | |
| 6 | 11 | 10/07 | Transfection imaging/ Make-Up | Midterm releases |
| 12 | 10/09 | Midterm: dedicated in class time | Midterm due Fri midnight | |
| 7 | 13 | 10/14 | CURE Intro | CURE Project Groups |
| 14 | 10/16 | CURE Begins | IF #1 Cell lines: PC3 (prostate cancer) DU145 “ LNCap “ MEF VPA (inc euchromtatin) MEF DZNep (dec heterochromatin) MEF LA KD (lamin A knockdown) HT1080 VPA and DZNEP Fix: 4% PFA + triton Block: PBS-Tw-Az-BSA for 15 min Primaries prepared by Drew’s lab: HK27ac MA5-23516 Thermo Fisher H3K9me2/3 Mouse CST 5327s H3K27me3 Rabbit CST 9733s H3K9ac Rabbit CST 9649s Secondaries: Rabbit green (enough for 50) Mouse red (enough for 50) | |
| 8 | 15 | 10/21 | CURE | IF #2 2nd batch of 18 Prostate cancer coverslips |
| 16 | 10/23 | CURE | Imaging & Analysis #1 | |
| 9 | 17 | 10/28 | CURE (Drew out) | Imaging & Analysis #2 |
| 18 | 10/30 | CURE | Imaging & Analysis #3 | |
| 10 | 11/04 | Holiday – Election Day | ||
| 19 | 11/06 | CURE | Graph | |
| 11 | 11/11 | Holiday – Veteran’s Day | ||
| 20 | 11/13 | CURE | CURE PPTX & report | |
| 12 | 21 | 11/18 | CURE last week – prepare report | CURE PPTX & report |
| 22 | 11/20 | CURE last week – prepare report | CURE PPTX & report | |
| 13 | 23 | 11/25 | CURE written report DUE | |
| 11/27 | THANKSGIVING: NO CLASS | |||
| 14 | 24 | 12/2 | CURE finalization | |
| 25 | 12/4 | CURE finalization | ||
| 15. | 26 | 12/9 | FLEXIBLE last day of class | |
| – | NO FINAL | THIS CLASS HAS NO FINAL | NO FINAL |