Name: finger-stick blood sampling methodology for the determination of exercise-induced lymphocyte apoptosis
Uploaded: 2011-02-24T17:30:00
Description: Watch this Scientific Journal Video about Finger-stick Blood Sampling Methodology for the Determination of Exercise-induced Lymphocyte Apoptosis at JoVE.com

This research was supported by a New Faculty Grant from the Faculty Scholarship Council at Western Kentucky University.

1. Finger-Stick Blood Sampling
<ol>
<li>Collection of whole blood is typically taken at rest for a baseline measurement (usually following 10-15 min seated rest), during or immediately following an exercise bout, and throughout the post-exercise period (generally from 1-3 hours following cessation of the exercise).</li>
<li>Using Universal Precautions, first sterilize the fingertip with 70% isopropyl alcohol.</li>
<li>Next, puncture the site using an automated lancet or similar device. Although we use a 30-gauge l...

<ol>
	<li>Simpson, R. J., Florida-James, G. D., Whyte, G. P., Black, J. R., Ross, J. A., Guy, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apoptosis+does+not+contribute+to+the+blood+lymphocytopenia+observed+after+intensive+and+downhill+treadmill+running+in+humans.">Apoptosis does not contribute to the blood lymphocytopenia observed after intensive and downhill treadmill running in humans.</a> Res. Sports Med. 15, 157-174 (2007).</li><li>Steensberg, A., Morrow, J., Toft, A. D., Bruunsgaard, H., Pedersen, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+exercise,+lymphocyte+apoptosis+and+F2-isoprostanes.">Prolonged exercise, lymphocyte apoptosis and F2-isoprostanes.</a> Eur. J. Appl. Physiol. 87, 38-42 (2002).</li><li>Mooren, F. C., Bloming, D., Lechtermann, A., Lerch, M. M., V&#246;lker, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymphocyte+apoptosis+after+exhaustive+and+moderate+exercise.">Lymphocyte apoptosis after exhaustive and moderate exercise.</a> J. Appl. Physiol. 93, 147-153 (2002).</li><li>Peters, E. M., Eden, M. V. a. n., Tyler, N., Ramautar, A., Chutergoon, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+exercise+does+not+cause+lymphocyte+DNA+damage+or+increased+apoptosis+in+well-trained+endurance+athletes.">Prolonged exercise does not cause lymphocyte DNA damage or increased apoptosis in well-trained endurance athletes.</a> Eur. J. Appl. Physiol. 98, 124-131 (2006).</li><li>Wang, J. -. S., Huang, Y. -. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+exercise+intensity+on+lymphocyte+apoptosis+induced+by+oxidative+stress+in+men.">Effects of exercise intensity on lymphocyte apoptosis induced by oxidative stress in men.</a> Eur. J. Appl. Physiol. 95, 290-291 (2005).</li><li>Mooren, F. C., Lechtermann, A., V&#246;lker, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise-induced+apoptosis+of+lymphocytes+depends+on+training+status.">Exercise-induced apoptosis of lymphocytes depends on training status.</a> Med. Sci. Sports Exerc. 36, 1476-1483 (2004).</li><li>Navalta, J. W., Sedlock, D. A., Park, K. -. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+treatment+influences+the+yield+of+apoptotic+lymphocytes+after+maximal+exercise.">Blood treatment influences the yield of apoptotic lymphocytes after maximal exercise.</a> Med. Sci. Sports Exerc. 37, 369-373 (2005).</li></ol>

A minimally invasive sample collection procedure and subsequent analysis is demonstrated for the analysis of both early and late phases of exercise-induced lymphocyte apoptosis. It is also possible to clearly exclude those cells, which are necrotic, and not apoptotic. This procedure overcomes the drawbacks and discomfort associated with invasive venipuncture from the antecubital region at multiple time points prior to, during, and immediately after exercise, or securing a catheter for the duration of the exercise bout. I...

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finger-stick blood sampling methodology for the determination of exercise-induced lymphocyte apoptosis

Exercise is a physiological stimulus capable of inducing apoptosis in immune cells. To date, various limitations have been identified with the measurement of this phenomenon, particularly relating to the amount of time required to isolate and treat a blood sample prior to the assessment of cell death. Because of this, it is difficult to determine whether reported increases in immune cell apoptosis can be contributed to the actual effect of exercise on the system, or are a reflection of the time and processing necessary to eventually obtain this measurement. In this article we demonstrate a rapid and minimally invasive procedure for the analysis of exercise-induced lymphocyte apoptosis. Unlike other techniques, whole blood is added to an antibody panel immediately upon obtaining a sample. Following the incubation period, red blood cells are lysed and samples are ready to be analyzed. The use of a finger-stick sampling procedure reduces the volume of blood required, and minimizes the discomfort to subjects.

Watch this Scientific Journal Video about Finger-stick Blood Sampling Methodology for the Determination of Exercise-induced Lymphocyte Apoptosis at JoVE.com

Finger-stick Blood Sampling Methodology for the Determination of Exercise-induced Lymphocyte Apoptosis

Exercise is capable of inducing apoptosis in immune cells. There are various measurement limitations, particularly relating to the amount of time required to isolate and treat a blood sample prior to the assessment. Demonstrated is a rapid and minimally invasive procedure for the analysis of exercise-induced lymphocyte apoptosis.

Exercise is a physiological stimulus capable of inducing apoptosis in immune cells. To date, various limitations have been identified with the measurement ...

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