Hosuk:LabNotes/2013-11-6: Difference between revisions

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>Hosuki78
(Created page with "*LabNote ===Movement test before after washing, stripping=== ====Procedure==== *Image - ACTB (ATTO550) *Aspirate, Add Formamide, Incubate, Aspirate, Was...")
 
>Mzcai
Line 28: Line 28:
*#Step2 : Strip
*#Step2 : Strip
*#Step3 : 1st Rolony + RAB7A
*#Step3 : 1st Rolony + RAB7A
*#Step4-0 : Strip
*#Step4 : RAB7A only
*#Step4 : RAB7A only
*#Step5-0 : Strip
*#Step5-0 : Strip
Line 35: Line 36:
*#Step7-0 : Strip
*#Step7-0 : Strip
*#Step7 : ACTB + RAB7A only
*#Step7 : ACTB + RAB7A only


====Result====
====Result====

Revision as of 00:06, 7 November 2013


Movement test before after washing, stripping

Procedure

  • Image - ACTB (ATTO550)
  • Aspirate, Add Formamide, Incubate, Aspirate, Washing with PBS
  • Image again - ACTB (ATTO550)


Result

  • 2 pixel off, 281.25nm x 2 = 562.5 nm off to the right direction

File:Exp1 MovementTest Before AfterWashing ACTB.png



ACTB, RAB7A probing test

  • 1st Rolony probe (Cy5) + ACTB (ATTO488) + RAB7A (ATTO550)  ACTB and RAB7A showed always weak signal.
  • Previous experiment (10/29/13) ACTB were seen more when added ACTB only.
  • So, let’s compare between RAB7A or ACTB and 1st Rolony probe(Cy5) together.
  • And compare to ACTB + RAB7A mix


Procedure

    1. Step1 : RAB7A only
    2. Step2 : Strip
    3. Step3 : 1st Rolony + RAB7A
    4. Step4-0 : Strip
    5. Step4 : RAB7A only
    6. Step5-0 : Strip
    7. Step5 : 1st Rolony + ACTB
    8. Step6-0 : Strip
    9. Step6 : ACTB only
    10. Step7-0 : Strip
    11. Step7 : ACTB + RAB7A only

Result

  • ACTB only had more than 1st Rolony+ACTB
  • RAB7A only had more than 1st Rolony+RAB7A
  • ACTB+RAB7A was similar to ACTB only or RAB7A only.


  • Step1 : RAB7A only,
    • RAB7A, Count = 514, Overlapped = 160


  • Step3 : 1st Rolony + RAB7A,
    • 1st Rolony Count = 5066
    • RAB7A, Count = 186, Overlapped = 114


  • Step4 : RAB7A only
    • RAB7A, Count = 591, Overlapped = 260


  • Step5 : 1st Rolony + ACTB
    • 1st Rolony Count = 5105
    • ACTB, Count = 137, Overlapped = 74


  • Step6 : ACTB only
    • ACTB, Count = 246, Overlapped = 157


  • Step7 : ACTB + RAB7A only
    • ACTB, Count = 234, Overlapped = 130
    • RAB7A, Count = 463, Overlapped = 268