ImmunofluorescentstainingofSeaUrchinembryos
1. Transfer fixed embryos to microfuge tubes. Allow to settle for 10 minutes.Gently remove most of the liquid.2. Add 100 ul antibody to one tube and 100 ul 1% normal goat serum to the other as a control.Let sit for 45 minutes at room temperature. Embryos will settle.3. Remove most of liquid. Add 1 ml SW to wash.Allow to settle, remove liquid. 4. Add 100 ul Fluorescein- or Texas red-congugated anti-mouse IgG(dilt......阅读全文
Immunofluorescent－staining－of－Sea－Urchin－embryos
1. Transfer fixed embryos to microfuge tubes. Allow to settle for 10 minutes.Gently remove most of the liquid.2. Add 100 ul antibody to one tube and 1
Histochemical－staining－of－sea－urchin－embryos－for－(AP)－enzyme－activity
Histochemical staining of sea urchin embryos for alkaline phosphatase (AP) enzyme activity1. Obtain embryo samples, tube of AP substrate buffer and tu
Effect－of－UV－Treatment－on－Early－Development－of－Sea－Urchin－Embryos
Objective:To test the effects of UV radiation at 254nm on Sea Urchin embryo development after radiation at the 2-cell stage.Background:Sea Urchins exh
Look－Ma,－No－Archenteron!－Sulfates－role－in－sea－urchin－early－development
ObjectiveTo observe the role sulfate plays in sea urchin gastrulation, and to replicate the findings of Karp and Solursh, that sea urchin embryos fail
Sea－Urchin－Fertilization
Gametes are collected as described. To remove some of their jelly coat,wash eggs several times in ASW by allowing them to settle and gently pouring of
The－Effects－of－NiCl2on－Spicule－Formation
The Effects of NiCl2on Spicule FormationJessica Ann Billet, Franklin and Marshall, Class of 2000Background and ObjectiveSea urchins exhibit radial hol
Solutions－for－Sea－Urchin－Development
Solutions for Sea UrchinASW: Artificial sea water (MBL)NaCl24.72 gKCl0.67 gCaCl2.2H2O1.36 gMgCl2.6H2O4.66 gMgSO4.7H2O6.29 gNaHCO30.18 gdH2Oto 1 literS
Sea－Urchin－staging－series
Invertebrate embryos develop at different rates depending on the temperature of incubation. Descriptive stages based on the number of cells and their
Separation－of－sea－urchin－blastomeres
Objective: to separate the Blastomere at different points in development and observe the growth of the resulting embryos.Procedure: We extractrf the g
Sea－Urchin－Gamete－collection
Spawning can be induced in adult sea urchinsby injecting 1 ml of a0.5M KCl solution into several sites in the soft membrane around the mouth. Within m
Sea－Urchin－Animal－Maintenance
The following is written to help non-marine biologists use sea urchins occasionally in teaching; disregard if you have running sea water tanks in your
The－Effects－of－Ultraviolet－Light－on－the－Fertilization
Jill K. Flemming, Franklin & Marshall College, Class of 2001IntroductionThe objective of this project is to observe the effects of UV radiation on bot
The－effect－of－lithium－chloride－on－Lytechinus－variegates－embryos
PurposeThis experiment will study effects of lithium chloride on sea urchin development, forcusing on archenteron formation.IntroductionGastrulation i
Wholemount－staining－of－embryos
Fix embryos in formalin or MEMFA for one hour at room temperature with mixing. Rinse with TBS, replace with methanol, store at -20oC.Rehydrate by slow
The－effect－of－temperature－on－the－embryonic－development－of－sea－urchins
Objective: This experiment aims to discover if temperature has an effect on the rates of sea urchin embryonic development. By altering the external en
The－Effects－of－CalciumFree－Seawater－onthe－Development－ofSea－Urchin－Embryos
ObjectiveThe purpose of this experiment is to determine the effects of calcium-free seawater on the development of sea urchin embryos. Embryos were pl
Preparation－of－fixed－embryos－for－immunocytochemistry－and－AP－staining
1. Transfer 50 ml of embryo cultures to centrifuge tubes. Spin at 1500 rpm for 5 minutes. Check that you can see a pellet of embryos at the bottom.Qui
Whole－mount－antibody－staining－ofzebrafish－embryos－formarkers－ofsegmentation
1. Dechorionate 26 hr embryos (pharyngula stage) carefully with two fine forceps.Transfer to fixative (1% formaldehyde in PBS). Fix for 1 hour rocking
The－effects－ofdifferent－concentrations－oflithium－chloride－on－thedevelopment
AbstractPrevious studies by Stachel and colleagues indicate that lithium chloride induction of post-midblastular Brachydanio rerio embryos results in
DEVELOPMENTAL－EFFECTS－OF－LITHIUM－CHLORIDE－ON－XENOPUS－EMBRYOS
PURPOSE: The objective of this experiment is to study the effect of lithium chloride treatment on early blastula xenopus embryos.INTRODUCTION: When th
Zebrafish－whole－mou...
实验概要Whole mount staining of Zebrafish embryos, now commonly used, requires extra steps to fix and permeabilize to ensure the egg membrane is perme
Fixation－of－Embryos
MEMFA Fix10xMEMFA Salts1 part 10x MEMFA salts1 M MOPS1 part 37% formaldehyde20mM EGTA8 parts water10mM MgSO410x salts can be autoclaved and stored. Tu
Chick－or－Mouse－embr...
实验概要The following procedure describes the procedure for whole mount staining of chick or mouse embryo’s. A similar procedure could be used for sta
Isolation－of－mouse－embryos
1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesome
Isolation－of－Zebrafish－embryos
Zebrafish will mate and deposit fertilized eggs on the bottom of the tank at 'dawn'. They can be accustomed to lay at any convenient time by k
Determining－the－effects－of－nicotine－on－bone－－in－chicks
ObjectiveThe purpose of this experiment is to determine how the development of bone and cartilage in a chick are affected by treating the embryo with
Protocol－to－Count－Cell－Number－of－Preimplantation－Embryos
Protocol to Count Cell Number of Preimplantation Embryos using Nuclear Staining with Hoechst 33342 or DAPI Introduction The following is a simple pro
Staining－Methods－for－cell
death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blueNon-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidiu
Protein－Staining－Procedures
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
Fluorescent－Staining－of－Cells
1. Fluorescent phalloidin in methanol. Phallacidin does not work as well. Dilute 10 ul 330 nM stock into 500 ul PBS for each large coverslip.2. PB