Announcements Please pick up syllabus and index card from front desk. –Fold the card lengthwise, write your name big and bold and use it as a name plate.

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Presentation transcript:

Announcements Please pick up syllabus and index card from front desk. –Fold the card lengthwise, write your name big and bold and use it as a name plate on your table. Lecture notes will be available on Blackboard before class. TBA this week: continue working with your samples, practicing image collection and file management. –For viewing, sign up on confocal room door and with either me or Phil Oshel.

Agenda, Week 1 A.Introduction to Confocal: Syllabus, etc. B.Lab: Fixation and staining of brine shrimp with fluorescent probes 1.Sytox Green for DNA 2.Rhodamine-phalloidin for microfilaments (enriched in muscle) 3.Unstained control for background fluorescence C.Seminar from 4-5, Brooks 176: Heather Wiatrowski, Microbiology job candidate. D.TBA this week and next: complete staining, view results with Dr. Hertzler or Phil Oshel.

TBA Availability with Dr. Hertzler: Two 3-student groups, 2 hours each TimeTuesdayWednesdayThursday 8 9SEMCell Biology 10Office 11Hours UCC 4Faculty MeetingSeminar

Artemia Staining Artemia at 1, 2, and 3 days (at 30 o C) are available. Fix with 3.7% formaldehyde in artificial seawater (ASW) for 1-2 hours. After seminar: Wash 3 X 5 min with ASW. Each group should label 3 tubes and stain overnight, covered with tin foil, with rocking: a)Sytox Green: 1:5000 in ASW (gloves) b)Rhodamine-phalloidin: 1:40 in ASW (gloves) c)nothing (control) Tuesday, in Microscope suite (one locker available per group): Wash 3 X 5 min with ASW. Mount with either: a)50% glycerol/50% ASW then 90% glycerol (store in fridg until TBA time), or b)EtOH dehydration, 3X in 100% EtOH, then methyl salicylate (store at RT in dark)

Introduction to Confocal Imaging A.Confocal versus conventional fluorescence B.Optical sectioning C.Imaging modes and applications D.Advantages, limitations of confocal

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Human brain sliceRabbit muscle fibers Sunflower pollen grain Widefield Confocal