Dec 07, 2023

Public workspaceLuminescent Conjugated Oligothiophenes (LCO) Staining

DOI
dx.doi.org/10.17504/protocols.io.x54v9pd7pg3e/v1
  • 1Department of Clinical and Movement Neurosciences, UCL Institute of Neurology, Queen Square, London WC1N 3BG, UK;
  • 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20 815, USA;
  • 3Queen Square Brain Bank for Neurological Disorders (QSBB), 1 Wakefield Street, London WC1N 1PJ, UK
Toby J Curless
Open access
DOI: dx.doi.org/10.17504/protocols.io.x54v9pd7pg3e/v1
Protocol Citation: Toby J Curless, Hemanth Ramesh Nelvagal, Zane Jaunmuktane 2023. Luminescent Conjugated Oligothiophenes (LCO) Staining. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9pd7pg3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 07, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 86046
Keywords: ASAPCRN, Luminescent Conjugated Oligothiophenes, Brain sections
Funders Acknowledgement:
The Michael J. Fox Foundation for Parkinson’s Research (MJFF) and the Aligning Science Across Parkinson’s (ASAP) Initiative
Grant ID: ASAP-000478
Abstract
The protocol describes Luminescent Conjugated Oligothiophenes staining on brain sections.
Materials
  • 100 % ethanol
  • Coverslips
  • DEPC H2O
  • Epifluorescence microscope (Leica) using FITC filters
  • Humidified chamber for 30 mins
  • HydromountTM aqueous non-fluorescing mounting media (National Diagnostics)
  • Hydrophobic ImmEdgeTM PAP pen (Vector laboratories)
  • PBS
  • pFTAA solution (diluted in PBS).
  • Xylene
Safety warnings
Attention
Breathing in xylene vapours in the air can cause irritation to the eyes, nose and throat and it can cause irritation, redness and swelling to the skin or eyes; and headaches, dizziness, vertigo and drowsiness. Please work to your institution approved health & safety policies, risk assessments and local procedures for the safe use, storage and disposal or xylene. Refer to the data safety sheet.
Preparation
Preparation
Generate tissue sections using standard microtome sectioning protocols.
Heat tissue dry tissue sections for Duration01:00:00 at Temperature60 °C
1h
Deparaffinisation and LCO Staining
Deparaffinisation and LCO Staining
45m

Safety information
Please refer to the Xylene Data Safety Sheet and follow local approved risk assessements and protocols for safe handling and disposal of xylene.



De-paraffinise sections by Duration00:05:00 washes in xylene (x2), Duration00:05:00 100 % ethanol (x2) and Duration00:05:00 in DEPC H2O (x1).

15m
Circle tissue on slide using a hydrophobic ImmEdgeTM PAP pen (Vector laboratories) to contain solution.
Prepare 1:500 pFTAA solution (diluted in PBS). All steps from here should be performed away from light.
Pipette pFTAA solution onto tissue and incubate at room temperature in a humidified chamber for Duration00:30:00 mins.

30m
Wash slides in PBS 3x.
Coverslip slides using HydromountTM aqueous non-fluorescing mounting media (National Diagnostics).