Malinow: Difference between revisions
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==Getting a Qdot into the cell== | ==Getting a Qdot into the cell== | ||
#Conjugate Qdot with secondary antibody f(ab) | #Conjugate [[Qdot]] with secondary antibody f(ab) | ||
#Incubate tissue with primary antibodies for [[AMPAR]] and [[PSD95]] | #Incubate tissue with primary antibodies for [[AMPAR]] and [[PSD95]] | ||
#Puff Qdots onto cell body, these will bind the primary at [[AMPAR]] N-terminus | #Puff Qdots onto cell body, these will bind the primary at [[AMPAR]] N-terminus | ||
#When AMPARs internalize the Qdot will be dragged into cell | #When AMPARs internalize the [[Qdot]] will be dragged into cell | ||
#Cleave N-terminus of [[AMPAR]] to liberate Qdot | #Cleave N-terminus of [[AMPAR]] to liberate [[Qdot]] | ||
#Qdot can then bind the primary ligated to [[PSD95]] | #[[Qdot]] can then bind the primary ligated to [[PSD95]] | ||
==Notes== | ==Notes== | ||
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* Acidic basic polypeptide recognition sequences | * Acidic basic polypeptide recognition sequences | ||
* Talk with nanotech group about various ways to conj. Qdots | * Talk with nanotech group about various ways to conj. Qdots | ||
* Nichol and England - couple Qdot to [[AMPAR]] agonist | * Nichol and England - couple [[Qdot]] to [[AMPAR]] agonist | ||
* Have simulation be a competitive model where AMPARs are competing during [[LTP]] | * Have simulation be a competitive model where AMPARs are competing during [[LTP]] | ||
* Quantitative review on synaptic numbers (Sheng) | * Quantitative review on synaptic numbers (Sheng) | ||
Revision as of 15:45, 3 July 2013
Getting a Qdot into the cell
- Conjugate Qdot with secondary antibody f(ab)
- Incubate tissue with primary antibodies for AMPAR and PSD95
- Puff Qdots onto cell body, these will bind the primary at AMPAR N-terminus
- When AMPARs internalize the Qdot will be dragged into cell
- Cleave N-terminus of AMPAR to liberate Qdot
- Qdot can then bind the primary ligated to PSD95
Notes
- FLASH technology
- Bredt
- minisog - gfp
- Acidic basic polypeptide recognition sequences
- Talk with nanotech group about various ways to conj. Qdots
- Nichol and England - couple Qdot to AMPAR agonist
- Have simulation be a competitive model where AMPARs are competing during LTP
- Quantitative review on synaptic numbers (Sheng)
PALM STORM
There are two major groups of methods for functional super-resolution microscopy:
1. Deterministic super-resolution: The most commonly used emitters in biological microscopy, fluorophores, show a nonlinear response to excitation, and this nonlinear response can be exploited to enhance resolution. These methods include STED, GSD, RESOLFT and SSIM.
2. Stochastical super-resolution PALM STORM: The chemical complexity of many molecular light sources gives them a complex temporal behaviour, which can be used to make several close-by fluorophores emit light at separate times and thereby become resolvable in time. These methods include SOFI and all single-molecule localization methods (SMLM) such as SPDM, SPDMphymod, PALM, FPALM, STORM and dSTORM.
NRSA
- Dominant negative PSD95