Dear Mayank Verma

I advise you to read this publication :

Eur J Clin Invest. 2002 Jun;32(6):443-7.

Procion orange tracer dye technique vs. identification of intrafibrillar fibronectin in the assessment of sarcolemmal damage.

Palacio J, Gāldiz JB, Alvarez FJ, Orozco-Levi M, Lloreta J, Gea J.

Source

Department of Pneumology and Pathology, Hospital Del Mar - IMIM, Universitat Pompeu Fabra, Barcelona, Spain.

Best regards

Fabio

Dear Mayank Verma,

another article, perhaps also of interest to you could be:

http://php.scripts.psu.edu/staff/e/b/ebl3/eblimages/NatureMed.pdf

Greelish et al, 1999: Stable restoration of the sarcoglycan complex in dystrophic muscle perfused with histamine and a recombinant adeno-associated viral vector

(using Procion Orange, and some hint on Evans blue, interesting topic and images...). Also there exists another, old reference, Moens P., Baatsen PHWW, Marechal G. Increased susceptibility of EDL muscles from mdx mice to damage induced by contractions with stretch. J.Muscle Res Cell Motil.14, 446-451,1993 using Procion red as bathing solution for demonstration of sarcolemmal damage. Best wishes and good luck, Wolfgang

Dear Susan,

I didn't find any valuable information in the URL you posted, dealing with the requested matter (sarcolemmal membrane damage localization):

the list retrievable as pdf consists of several dye combinations (with ProcionMX) to achieve special color tones.

The basic information on (used in that particular application more/less as cloth fibre stain) can be found on

http://www.jacquardproducts.com/procion-mx.html

and instructions for use are available @http://www.jacquardproducts.com/assets/jacquard-site/product-pages/dyes/procion-mx/Procion%20MX%20Instructions.pdf.

One for sure could try whether the coloring dye intended for fabrics or used as dye for artists is suited for the task of localizing sarcolemmal (muscle fibre) membrane defects, but there is not any hint on such incorporated in the information leaflets on JACQUARD's website (http://www.jacquardproducts.com/company/) . Disclaimer: no financial interest, no affiliation to that company.

Regards,

Wolfgang

One of my colleagues gave me the following protocol.

Procion Orange Staining

Solution

1% POD in PBS or Kreb Ringer buffer

Isolated muscle Protocol

1. Perform eccentric contraction in isolated muscle.

2. Incubate muscle is 1% POD in Kreb Ringer solution for 15 minute.

3. Freeze sample in liquid Nitrogen chilled isopentane in OCT

4. Cryosection at 10um.

5. Visualize at 580nm.

Infusion Protocol

100ul of 1% procion Orange in PBS can be injected IV by tail vein. The staining can be visualized between 1hr and 5 days. The paper decided to use 24 hour as their time point. (Covert persistence of mdx mouse myopathy is revealed by acute and chronic effects of irradiation.1999)

Good Morning,

dear Mayank, thank you very much for posting the protocol.

Just in case anybody could / would be willing to share the paper in this ResearchGate-Forum, I add here the reference data for the paper

(cf. PubMed: http://www.ncbi.nlm.nih.gov/pubmed/10402020) :

cf: http://www.sciencedirect.com/science/article/pii/S0022510X99000611,

Journal of the Neurological Sciences, Volume 164, Issue 2, 1 April 1999, Pages 103–116

C.N Pagel, T.A Partridge

< Covert persistence of mdx mouse myopathy is revealed by acute and chronic effects of irradiation >

Unfortunately for most of us this article is accessible only by PPV (pay per view).

It would - by the way - really helpful to know a little bit more about the Krebs-Ringer (a bicarbonate buffer) solution specifically used here for muscle fibres (knowing that there are some different Ringer solutions out there),

POD standing for Procion Orange Dye, I guess.

As another option I would like to point to the first paper mentioned by Fabio NOBILI in this posting thread:

J Palacio - ‎2002, where the following references are listed for:

5 Zhu E, Petrof BJ, Gea J, Comtois N, Grassino AE.

Diaphragm muscle fibre injury after inspiratory resistive breathing.

Am J Respir Crit Care Med 1997;155:1110–6.

6 Lin M Ch, Ebihara S, El Dwairi Q, Hussain SNA, Yang L, Gottfried SB et al. Diaphragm sarcolemmal injury is induced by sepsis and alleviated by nitric oxide synthase inhibition. Am J Respir Crit Care Med 1998;158:1656–63.

7 Petrof BJ, Shrager JB, Stedman HH, Kelly AM, Sweeney HL.

Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc Natl Acad Sci USA 1993;90:3710–4.

8 Ebihara S, Hussain SNA, Danialou G, Cho WK, Gottfried SB, Petrof BJ.

Mechanical ventilation protects against diaphragm injury in sepsis.

Am J Respir Crit Care Med 2002;165:221–8.

9 Paul GL, DeLany JP, Snook JT, Seifert JG, Kirby TE.

Serum and urinary markers of skeletal muscle tissue damage after weight lifting exercise. Eur J Appl Physiol Occup Physiol 1989;58:786–90.

10 Friden J, Sfakianos PN, Hargens AR.

Blood indices of muscle injury associated with eccentric muscle contractions.

J Orthop Res 1989;7:142–5.

11 Soritcher S, Mair J, Koller A, Gebert W, Rama D, Calzolari C et al.

Skeletal troponin I as a marker of exerciseinduced muscle damage.

J Appl Physiol 1997;83:1076–82.

12 Koller A, Mair J, Schobersberger W, Wohlfarter T, Haid C, Mayr M et al.

Effects of prolonged strenuous endurance exercise on plasma myosin heavy chain fragments and other muscular proteins. Cycling vs running.

J Sports Med Phys Fitness 1998;38:10–7.

13 Friden J, Lieber RL. Segmental muscle fiber lesions after repetitive eccentric contractions. Cell Tissue Res 1998;293:165–71.

14 Margaritis I, Tessier F, Verdera F, Bermon S, Marconnet P.

Muscle enzyme release does not predict muscle function impairment after triathlon. J Sports Med Phys Fitness 1999;39:133–9.

15 Sorichter S, Puschendorf B, Mair J. Skeletal muscle injury induced by eccentric muscle action: muscle proteins as markers of muscle fiber injury. Exerc Immunol Rev 1999;5:5–21.

For perhaps an (old) application alternative as fluorescent dye (extra-intracell.stain, the latter perhaps after membrane damage) on nerve fibers cf.

http://web.as.uky.edu/Biology/faculty/cooper/Bio450-AS300/Earth%20worm%20experiment/HISTOLOGICAL%20STAINING%20PROPERTIES.pdf

PAYTON BW (1970), Histological Staining properties of Procion Yellow M4RS

(= J Cell Biol. 1970 Jun 1;45(3):659–662) in which the author argues that intraaxonal staining with the dye perhaps is/was due to "open" ends of sectioned axons providing the possibility to enter the axonal body cut at their ends lying in the dye bath.

Best wishes and regards, Wolfgang