Lab outcomes – May 18-22

Mon

Setup spawning tanks
Water change in all medium tanks & glass bowl

Tues

Spawning: ~400 embryos; used in PH#20
No microinjection
Breakdown/setup tanks
RT-PCR with June

Wed

Spawning: ~150 embryos; used in PH#20
No microinjection
Checked paramecia
Breakdown/setup tanks
RT-PCR with June

Thurs

Spawning: ~170 embryos; used in PH#20
Worked at JARTU with Ann

Fri

Made paramecia boxes
Meeting

Schedules – May 4-8

Jack
June
Emily
Ontology analysis
Ping
Regular work – fish colony maintenance & spawning
Ann
Exam week

Candace
Rachel & DeOnna & Rachel D.
Exam week

Schedules – April 20-24

Jack
?????
June
Emily
Ontology analysis
Ping
Mon – setup spawning; water change in medium tanks; take care of larvae
Tues – collect embryos; break down/setup spawning
Wed – collect embryos; break down spawning/setup spawning
Thurs – break down spawning with Ann
Fri – work on website; water quality tests
Ann
Mon
Check paramecia ~30min
Make BG-11 media ~1hr
Take pics and clean eggs ~1hr
Tues
Centrifuge microcystis ~7hr
Practice weighing and [...]

Schedule – week of March 2

Jack
Getting the YFP construct going
June
Emily
M-F 9am-5pm
Pathway analysis
Reproduction experiment
Ping
Mon – setup spawning; water change in medium tanks; brine shrimp acid treatment experiment; larvae care
Tues – collect embryos; break down/setup spawning; brine shrimp acid treatment experiment
Wed – collect embryos; break down / setup Ann’s spawning tanks; reproduction exp.; brine shrimp acid treatment experiment
Thurs – reproduction exp.; brine [...]

Lab outcomes week Jan 19-23

Mon

MLK Day

Tues

Water change in medium tanks & big tanks

Wed

RNA extraction
Setup spawning tanks
Jack & Ping met with Jill

Thurs

Spawning: ~200 embryos; all discarded
No microinjection – Candace was sick
Breakdown tanks; setup tanks
RNA extraction
Cytotiva observation
Paramecia checked

Fri

Spawning: ~ 500 embryos; labeled 012309
Microinjection – ~250 embryos
Histology
Cytoviva observation
Rachel D. doing histology with DeOnna and Rachel J.

Lab outcomes week Nov 24-28

Mon

Setup spawning tanks
All October larvae doing well
Dissection practice – June & Ping
Separated males (44) and females (64) in Jan/Feb batch
Microcystis & EE2 experiment started (Ann’s)

Tues

Spawning: ~500 embryos; discarded
No microinjection
Water change in medium tank and bowl

Wed

Dissection practice
Paramecia split
Separated males (64) & females (68) in March-July batch

Thurs

Thanksgiving holiday
Changed water for larvae

Fri

Thanksgiving holiday
Ending Ann’s experiment

Lab outcomes – Nov 10-14

Mon

Setup spawning tanks
Larvae 102108 Day 20 & 102308 Day 18 – BS; growing well
102808 Day 13 – BS; growing well
103008 Day 11 – BS; observed some orange bellies
Discarded JWP P#1 – mass dying in exposure beakers
Setup new JWP P#1 @ 50% of original concentration

Tues

Spawning: ~150 embryos; all kept and labeled as 111108
Microinjection ~50 embryos
Fish room [...]

Microinjection Results

Candace Schaefer
Microinjection Report: 10/16/08
Submitted 10/20/08

1. Set-up

1.5% agar slants
Vortexed C60 soln 30 sec

C60 soln = 09/09/08 supernatant from 10 mg/ml C60 in DMSO

Centrifuged soln 15 sec
5% green C60

95 ul C60-DMSO
5 ul green food dye

Centrifuged 5% green C60 and DMSO (control) 10 min @ 10 rpm
Loaded needles
Set psi = 30 and broke needles

[...]

Candace’s microinjection results

DiI:DMSO (1:4)
 

well

20-50 ms

100 ms

200 ms

500 ms

F1

F2

 

 

 

 

F3

 

72 hpf
(same fish 3 images below)
 
 

F4

 

 

 

F5

 

 

72 hpf
(malformed)

F6

 

 

72 hpf
 

F8

 

 

 

72 hpf

500 ms
 
          

 
 

Candace’s microinjection results

Candace Schaefer
Solutions 09/12/08
DiI-EtOH in PBS (1:1)
50 ms

DiI-EtOH in PBS (1:1)

DiI-EtOH in DMSO
1:1?
5ms                     5 ms (bright)
1:2?
25 ms                    5 ms (bright)
1:4
25 ms
1:9
25 ms                25 ms (bright)            25 ms (bright)
DiI-EtOH in C60-DMSO
1:4
25 ms                25 ms                      25 ms        [...]