Name: Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique (Video) | JoVE Journal
Uploaded: 2022-09-13T15:00:00.000+00:00
Duration: 4 min 52 s
Description: Learn a non-invasive microdialysis technique for dynamic continuous blood extraction from rat hearts. This method enables real-time cardiovascular biomarker analysis without sample pre-processing.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

This work was supported by the National Natural Science Foundation of China (82104533), the China Postdoctoral Science Foundation (2020M683273), the Science &#38; Technology Department of Sichuan province (2021YJ0175) and the Key R&#38;D project of Sichuan Provincial Science and Technology Plan (2022YFS0438). Meanwhile, the authors would like to thank Mr. Yuncheng Hong, a senior equipment engineer at TRI-ANGELS D&#38;H TRADING PTE. LTD. (Singapore city, Singapore), for providing technical services for microdialysis techniques.

The animal protocol has been approved by the Administrative Committee of Chengdu University of Traditional Chinese Medicine (Record number: 2021-11). Specified pathogen-free male Sprague Dawley (SD) rats (8-10 weeks, 260-300 g) were raised in independent ventilation cages, maintaining the laboratory environment at 22 &#176;C and 65% relative humidity, and were used for the present study. The animals were obtained from a commercial source (see Table of Materials). All rats were habituated to adaptive feeding for 1 week with free water and diet during the period.
1. Experimental preparation
<ol>
	<li>Assemble the equipment involved in microdialysis of blood sampling (see Table of Materials), as shown in Figure 1.</li>
	<li>Prepare anti-coagulant citrate dextrose solution (ACDs), containing 3.50 mmol/L of citrate, 7.50 mmol/L of sodium citrate, and 13.60 mmol/L of glucose, as the perfusion fluid for microdialysis blood collection (see Table of Materials).</li>
	<li>Before use, filter the ACDs with a 0.22 &#181;m membrane filtration unit and remove bubbles ultrasonically. Keep ACDs at 37 &#176;C to reduce the stimulation for SD rats.</li>
</ol>
2. Patency inspection of microdialysis piping system
<ol>
	<li>Attach the inlet of the probe of the dialysis unit with the syringe needle, the tubing adapter, and the fep tubing (see Table of Materials). 
	NOTE: The blue end of the microdialysis probe is for the fluid influx, and the transparent end is for the fluid outflow.</li>
	<li>Check the patency of the microdialysis piping system by perfusing ACDs18 into the piping system at a speed of 2 &#181;L/min. 
	&#8203;NOTE: If ACDs flow from the sample collection site, the unimpeded microdialysis system can be used for further cardiac blood collection. If not, the blood probe must be checked for broken leaks or whether the pipe-pipe joint was sealed.</li>
</ol>
3. Microdialysis probe implantation
<ol>
	<li>Anesthetize the rats with 2% isoflurane in an air-oxygen mixture of 0.6 L/min, fasten the thoroughly unconscious rats on the operating table, and maintain the body temperature at 37 &#176;C using an animal temperature maintainer (see Table of Materials).</li>
	<li>Remove the fur from the neck with an electric shaver and disinfect the surgical site with three alternating rounds of betadine and 70% alcohol. Inject bupivacaine (1.5 mg/kg) into the rat. Expose the right jugular vein by blunt dissection of soft tissue and perivascular fascia through a 1.5 cm incision along the midline of the neck. 
	NOTE: All surgical instruments and tools used in the experiment should be sterilized in advance by autoclaving. Use sterile gloves and surgical drapes. The whole experimental operation should be carried out in a sterile environment. Follow local animal use guidelines for anesthesia and analgesia regimen.</li>
	<li>Make a detachable slipknot using a 4-0 surgical suture in the right jugular vein at the distal end of the heart to temporarily block blood flow, and make a 1.5 cm incision in the right jugular vein near the heart.</li>
	<li>Insert a needle-shaped catheter stylet (length 25 mm, diameter 0.7 mm, see Table of Materials) into the right jugular vein toward the proximal end of the rat heart, insert the blood microdialysis probe (total probe length 24 mm, membrane length 10 mm) in a catheter, and implant the probe with ophthalmic forceps along the oblique incision of the catheter stylet19.</li>
	<li>Remove the guided catheter stylet and fully immerse the semi-permeable membrane of the probe (membrane length 10 mm, membrane diameter 0.5 mm) in the right jugular vein. Unravel the detachable slipknot at the distal end of the heart to restore blood flow in the right jugular vein.</li>
	<li>Ligature the probe with the right jugular vein using 4-0 surgical sutures and subcutaneously thread the probe tail tube through the back of the neck. See Figure 2 for specific probe implantation steps. 
	&#8203;NOTE: The molecular cut-off by the blood microdialysis probe used in this study is &#62;20 kD. The collected blood samples contain substances with a molecular weight of less than 20 kD.</li>
</ol>
4. Microdialysis sampling
<ol>
	<li>A week after the microdialysis probe implantation, rats recovering from surgical trauma undergo microdialysis sampling. Place the awake rat in a free-moving tank (see Table of Materials) and connect the probe to the microdialysis system. Equilibrate the probe dialysis membrane by irrigating ACDs with a rate of 2 &#181;L/min for 1 h.</li>
	<li>Collect microdialysis blood samples at a flow rate of 2 &#181;L/min and temporarily keep them in a 4 &#176;C fractional container. 
	&#8203;NOTE: The obtained blood samples could be tested directly by centrifuging at 20,000 x g for 10 min at room temperature. Or it could be stored at -80 &#176;C for the following testing. When collecting samples, it is important always to observe whether the probe prolapse and/or leak out.</li>
</ol>
5. Post-sampling operation
<ol>
	<li>Anesthetize the SD rats with 2% isoflurane in an air-oxygen mixture at 0.6 L/min.</li>
	<li>Dissect to re-confirm that the probe is in the right jugular vein. Remove the microdialysis probe from the right jugular vein and put it into ultrapure water.</li>
	<li>Finally, euthanize the rats by inhalation of 5% isoflurane.</li>
	<li>Connect the probe to the pipeline and rinse overnight at a rate of 2 &#181;L/min with ultrapure water to completely wash out the residual salt in the pipe and probe.</li>
	<li>Remove the probe and soak it in ultrapure water. Store at 4 &#176;C to prevent the probe dialysis membrane from contracting. 
	NOTE: If the volume of dialysis fluid is inconsistent with the volume of perfusion fluid, the probe may be blocked by coagulated blood. The probe can be placed in the pancreatic protein solution until the visible substance flakes off from the tip of the probe membrane.</li>
</ol>

<ol><li>van Rensburg, W. J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Post-mortem+evidence+of+a+diverse+distribution+pattern+of+atherosclerosis+in+the+South+African+population%5BTitle%5D)+AND+%22Scientific+Reports%22%5BJournal%5D)">Post-mortem evidence of a diverse distribution pattern of atherosclerosis in the South African population.</a> Scientific Reports. 12 (1), 11366(2022).</li>
<li>Katz, A. J., Chen, R. C., Usinger, D. S., Danus, S. M., Zullig, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cardiovascular+disease+prevention+and+management+of+pre-existent+cardiovascular+disease+in+a+cohort+of+prostate+cancer+survivors%5BTitle%5D)+AND+%22Journal+of+Cancer+Survivorship%22%5BJournal%5D)">Cardiovascular disease prevention and management of pre-existent cardiovascular disease in a cohort of prostate cancer survivors.</a> Journal of Cancer Survivorship. , (2022).</li>
<li>Rødevand, L., Tesli, M., Andreassen, O. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cardiovascular+disease+risk+in+people+with+severe+mental+disorders%3A+an+update+and+call+for+action%5BTitle%5D)+AND+%22Current+Opinion+in+Psychiatry%22%5BJournal%5D)">Cardiovascular disease risk in people with severe mental disorders: an update and call for action.</a> Current Opinion in Psychiatry. 35 (4), 277-284 (2022).</li>
<li>Izumi, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Impact+of+circulating+cathepsin+K+on+the+coronary+calcification+and+the+clinical+outcome+in+chronic+kidney+disease+patients%5BTitle%5D)+AND+%22Heart+and+Vessels%22%5BJournal%5D)">Impact of circulating cathepsin K on the coronary calcification and the clinical outcome in chronic kidney disease patients.</a> Heart and Vessels. 31 (1), 6-14 (2016).</li>
<li>Wang, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Whey+protein+hydrolysate+alleviated+atherosclerosis+and+hepatic+steatosis+by+regulating+lipid+metabolism+in+apoE-%2F-+mice+fed+a+Western+diet%5BTitle%5D)+AND+%22Food+Research+International%22%5BJournal%5D)">Whey protein hydrolysate alleviated atherosclerosis and hepatic steatosis by regulating lipid metabolism in apoE-/- mice fed a Western diet.</a> Food Research International. 157, 111419(2022).</li>
<li>Angelone, T., Rocca, C., Pasqua, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nesfatin-1+in+cardiovascular+orchestration%3A+From+bench+to+bedside%5BTitle%5D)+AND+%22Pharmacological+Research%22%5BJournal%5D)">Nesfatin-1 in cardiovascular orchestration: From bench to bedside.</a> Pharmacological Research. 156, 104766(2020).</li>
<li>Bernardi, P. M., Barreto, F., Dalla Costa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Application+of+a+LC-MS%2FMS+method+for+evaluating+lung+penetration+of+tobramycin+in+rats+by+microdialysis%5BTitle%5D)+AND+%22Journal+of+Pharmaceutical+and+Biomedical+Analysis%22%5BJournal%5D)">Application of a LC-MS/MS method for evaluating lung penetration of tobramycin in rats by microdialysis.</a> Journal of Pharmaceutical and Biomedical Analysis. 134, 340-345 (2017).</li>
<li>Anderzhanova, E., Wotjak, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Brain+microdialysis+and+its+applications+in+experimental+neurochemistry%5BTitle%5D)+AND+%22Cell+and+Tissue+Research%22%5BJournal%5D)">Brain microdialysis and its applications in experimental neurochemistry.</a> Cell and Tissue Research. 354 (1), 27-39 (2013).</li>
<li>Joukhadar, C., Müller, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis%3A+current+applications+in+clinical+pharmacokinetic+studies+and+its+potential+role+in+the+future%5BTitle%5D)+AND+%22Clinical+Pharmacokinetics%22%5BJournal%5D)">Microdialysis: current applications in clinical pharmacokinetic studies and its potential role in the future.</a> Clinical Pharmacokinetics. 44 (9), 895-913 (2005).</li>
<li>Stangler, L. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis+and+microperfusion+electrodes+in+neurologic+disease+monitoring%5BTitle%5D)+AND+%22Fluids+and+Barriers+of+the+CNS%22%5BJournal%5D)">Microdialysis and microperfusion electrodes in neurologic disease monitoring.</a> Fluids and Barriers of the CNS. 18 (1), 52(2021).</li>
<li>Young, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cerebral+microdialysis%5BTitle%5D)+AND+%22Critical+Care+Nursing+Clinics+of+North+America%22%5BJournal%5D)">Cerebral microdialysis.</a> Critical Care Nursing Clinics of North America. 28 (1), 109-124 (2016).</li>
<li>O'Connell, M. T., Krejci, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis+techniques+and+microdialysis-based+patient-near+diagnostics%5BTitle%5D)+AND+%22Analytical+and+Bioanalytical+Chemistry%22%5BJournal%5D)">Microdialysis techniques and microdialysis-based patient-near diagnostics.</a> Analytical and Bioanalytical Chemistry. 414 (10), 3165-3175 (2022).</li>
<li>Hammarlund-Udenaes, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis+as+an+important+technique+in+systems+pharmacology-a+historical+and+methodological+review%5BTitle%5D)+AND+%22The+AAPS+Journal%22%5BJournal%5D)">Microdialysis as an important technique in systems pharmacology-a historical and methodological review.</a> The AAPS Journal. 19 (5), 1294-1303 (2017).</li>
<li>Stahl, M., Bouw, R., Jackson, A., Pay, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+microdialysis%5BTitle%5D)+AND+%22Current+Pharmaceutical+Biotechnology%22%5BJournal%5D)">Human microdialysis.</a> Current Pharmaceutical Biotechnology. 3 (2), 165-178 (2002).</li>
<li>Pierce, C. F., Kwasnicki, A., Lakka, S. S., Engelhard, H. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cerebral+microdialysis+as+a+tool+for+assessing+the+delivery+of+chemotherapy+in+brain+tumor+patients%5BTitle%5D)+AND+%22World+Neurosurgery%22%5BJournal%5D)">Cerebral microdialysis as a tool for assessing the delivery of chemotherapy in brain tumor patients.</a> World Neurosurgery. 145, 187-196 (2021).</li>
<li>Sørensen, M., Jacobsen, S., Petersen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis+in+equine+research%3A+a+review+of+clinical+and+experimental+findings%5BTitle%5D)+AND+%22Veterinary+Journal%22%5BJournal%5D)">Microdialysis in equine research: a review of clinical and experimental findings.</a> Veterinary Journal. 197 (3), 553-559 (2013).</li>
<li>Dmitrieva, N., Rodríguez-Malaver, A. J., Pérez, J., Hernández, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differential+release+of+neurotransmitters+from+superficial+and+deep+layers+of+the+dorsal+horn+in+response+to+acute+noxious+stimulation+and+inflammation+of+the+rat+paw%5BTitle%5D)+AND+%22European+Journal+of+Pain%22%5BJournal%5D)">Differential release of neurotransmitters from superficial and deep layers of the dorsal horn in response to acute noxious stimulation and inflammation of the rat paw.</a> European Journal of Pain. 8 (3), 245-252 (2004).</li>
<li>Li, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis+sampling+and+HPLC-MS%2FMS+quantification+of+sinomenine%2C+ligustrazine%2C+gabapentin%2C+paracetamol%2C+pregabalin+and+amitriptyline+in+rat+blood+and+brain+extracellular+fluid%5BTitle%5D)+AND+%22Acta+Pharmaceutica+Sinica%22%5BJournal%5D)">Microdialysis sampling and HPLC-MS/MS quantification of sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin and amitriptyline in rat blood and brain extracellular fluid.</a> Acta Pharmaceutica Sinica. 55 (9), 2198-2206 (2020).</li>
<li>Chauzy, A., Lamarche, I., Adier, C., Couet, W., Marchand, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis+study+of+Aztreonam-Avibactam+distribution+in+peritoneal+fluid+and+muscle+of+rats+with+or+without+experimental+peritonitis%5BTitle%5D)+AND+%22Antimicrobial+Agents+and+Chemotherapy%22%5BJournal%5D)">Microdialysis study of Aztreonam-Avibactam distribution in peritoneal fluid and muscle of rats with or without experimental peritonitis.</a> Antimicrobial Agents and Chemotherapy. 62 (10), 01228(2018).</li>
<li>Fang, X. X., Ardehali, H., Min, J. X., Wang, F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+molecular+and+metabolic+landscape+of+iron+and+ferroptosis+in+cardiovascular+disease%5BTitle%5D)+AND+%22Nature+Reviews.+Cardiology%22%5BJournal%5D)">The molecular and metabolic landscape of iron and ferroptosis in cardiovascular disease.</a> Nature Reviews. Cardiology. , 1-17 (2022).</li>
<li>Samson, R., Ennezat, P. V., Le Jemtel, T. H., Oparil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cardiovascular+disease+risk+reduction+and+body+mass+index%5BTitle%5D)+AND+%22Current+Hypertension+Reports%22%5BJournal%5D)">Cardiovascular disease risk reduction and body mass index.</a> Current Hypertension Reports. , (2022).</li>
<li>Kim, M. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((School+racial+segregation+and+long-term+cardiovascular+health+among+Black+adults+in+the+US%3A+A+quasi-experimental+study%5BTitle%5D)+AND+%22PLoS+Medicine%22%5BJournal%5D)">School racial segregation and long-term cardiovascular health among Black adults in the US: A quasi-experimental study.</a> PLoS Medicine. 19 (6), 1004031(2022).</li>
<li>Qin, Y. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Role+of+m6A+RNA+methylation+in+cardiovascular+disease+%28Review%29%5BTitle%5D)+AND+%22International+Journal+of+Molecular+Medicine%22%5BJournal%5D)">Role of m6A RNA methylation in cardiovascular disease (Review).</a> International Journal of Molecular Medicine. 46 (6), 1958-1972 (2020).</li>
<li>Xu, C. M., Liu, C. J., Xiong, J. H., Yu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cardiovascular+aspects+of+the+%28pro%29renin+receptor%3A+Function+and+significance%5BTitle%5D)+AND+%22FASEB+Journal%22%5BJournal%5D)">Cardiovascular aspects of the (pro)renin receptor: Function and significance.</a> FASEB Journal. 36 (4), 22237(2022).</li>
<li>Guvenc-Bayram, G., Yalcin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+intermediary+role+of+the+central+cyclooxygenase+%2F+lipoxygenase+enzymes+in+intracerebroventricular+injected+nesfatin-1-evoked+cardiovascular+effects+in+rats%5BTitle%5D)+AND+%22Neuroscience+Letters%22%5BJournal%5D)">The intermediary role of the central cyclooxygenase / lipoxygenase enzymes in intracerebroventricular injected nesfatin-1-evoked cardiovascular effects in rats.</a> Neuroscience Letters. 756, 135961(2021).</li>
<li>Ahrens Kress, A. P., Zhang, Y. D., Kaiser-Vry, A. R., Sauer, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+comparison+of+blood+collection+techniques+in+mice+and+their+effects+on+welfare%5BTitle%5D)+AND+%22Journal+of+the+American+Association+for+Laboratory+Animal+Science%22%5BJournal%5D)">A comparison of blood collection techniques in mice and their effects on welfare.</a> Journal of the American Association for Laboratory Animal Science. 61 (3), 287-295 (2022).</li>
<li>Joshi, A., Patel, H., Joshi, A., Stagni, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pharmacokinetic+applications+of+cutaneous+microdialysis%3A+Continuous%2Bintermittent+vs+continuous-only+sampling%5BTitle%5D)+AND+%22Journal+of+Pharmacological+and+Toxicological+Methods%22%5BJournal%5D)">Pharmacokinetic applications of cutaneous microdialysis: Continuous+intermittent vs continuous-only sampling.</a> Journal of Pharmacological and Toxicological Methods. 83, 16-20 (2017).</li>
<li>Reyes-Garcés, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((In+vivo+brain+sampling+using+a+microextraction+probe+reveals+metabolic+changes+in+rodents+after+deep+brain+stimulation%5BTitle%5D)+AND+%22Analytical+Chemistry%22%5BJournal%5D)">In vivo brain sampling using a microextraction probe reveals metabolic changes in rodents after deep brain stimulation.</a> Analytical Chemistry. 91 (15), 9875-9884 (2019).</li>
<li>Kho, C. M., Enche Ab Rahim, S. K., Ahmad, Z. A., Abdullah, N. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+review+on+microdialysis+calibration+methods%3A+the+theory+and+current+related+efforts%5BTitle%5D)+AND+%22Molecular+Neurobiology%22%5BJournal%5D)">A review on microdialysis calibration methods: the theory and current related efforts.</a> Molecular Neurobiology. 54 (5), 3506-3527 (2017).</li>
<li>Zhuang, L. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Theory+and+application+of+microdialysis+in+pharmacokinetic+studies%5BTitle%5D)+AND+%22Current+Drug+Metabolism%22%5BJournal%5D)">Theory and application of microdialysis in pharmacokinetic studies.</a> Current Drug Metabolism. 16 (10), 919-931 (2015).</li>
<li>Zhang, Y. F., Huang, X. X., Zhu, L. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Metabonomics+research+strategy+based+on+microdialysis+technique%5BTitle%5D)+AND+%22China+Journal+of+Chinese+Materia+Medica%22%5BJournal%5D)">Metabonomics research strategy based on microdialysis technique.</a> China Journal of Chinese Materia Medica. 45 (1), 214-220 (2020).</li>
<li>Carpenter, K. L., Young, A. M., Hutchinson, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Advanced+monitoring+in+traumatic+brain+injury%3A+microdialysis%5BTitle%5D)+AND+%22Current+Opinion+in+Critical+Care%22%5BJournal%5D)">Advanced monitoring in traumatic brain injury: microdialysis.</a> Current Opinion in Critical Care. 23 (2), 103-109 (2017).</li>
<li>Brunner, M., Langer, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microdialysis+versus+other+techniques+for+the+clinical+assessment+of+in+vivo+tissue+drug+distribution%5BTitle%5D)+AND+%22The+AAPS+Journal%22%5BJournal%5D)">Microdialysis versus other techniques for the clinical assessment of in vivo tissue drug distribution.</a> The AAPS Journal. 8 (2), 263-271 (2006).</li>
<li>Tettey-Amlalo, R. N., Kanfer, I., Skinner, M. F., Benfeldt, E., Verbeeck, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Application+of+dermal+microdialysis+for+the+evaluation+of+bioequivalence+of+a+ketoprofen+topical+gel%5BTitle%5D)+AND+%22European+Journal+of+Pharmaceutical+Sciences%22%5BJournal%5D)">Application of dermal microdialysis for the evaluation of bioequivalence of a ketoprofen topical gel.</a> European Journal of Pharmaceutical Sciences. 36 (2-3), 219-225 (2009).</li>
<li>Dhanani, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Recovery+rates+of+combination+antibiotic+therapy+using+in+vitro+microdialysis+simulating+in+vivo+conditions%5BTitle%5D)+AND+%22Journal+of+Pharmaceutical+Analysis%22%5BJournal%5D)">Recovery rates of combination antibiotic therapy using in vitro microdialysis simulating in vivo conditions.</a> Journal of Pharmaceutical Analysis. 8 (6), 407-412 (2018).</li>
</ol>

The authors have nothing to disclose.

CVDs are a common chronic disease in clinics with gradually increasing incidence in China, and the onset age tends to be younger, causing the concern and panic of most patients20,21. Being the leading cause of death in the world, CVDs can induce cerebral infarction and other high mortality diseases, seriously threatening the healthy life of patients22. CVDs, including ischemic heart disease, cardiomyopathy, atherosclerosis, high blood pres...

With the acceleration of the pace of life and the increase of psychological pressure, cardiovascular diseases (CVDs) tend to occur in young, middle-aged, and elderly people1,2. The morbidity and mortality of CVDs are high, with the characteristics of acute onset, rapid progression, and a long course of the disease, which seriously affect the safety of patients3. The occurrence of CVDs may be closely related to the changes in some blood components, such as cholesterol, serum lipids, blood glucose, myocardial enzymes, and protein kinase K4,5,6. The patient&#39;s relevant situation can be managed most quickly by analyzing routine blood examination items. Hence, the quality of the blood samples determines the accuracy of the test results. However, conventional methods for blood collection have some inevitable drawbacks, which seriously affect the experimental results, such as large trauma area, small blood collection volume, high requirements for operators, inability to reflect drug changes in real-time, cumbersome blood sample pretreatment, large consumption of experimental animals, and failure to meet animal ethical requirements7,8,9. With continuous advances in medical technology, the quality of blood collection has also put forward higher requirements. Therefore, it is urgent to develop a new blood sampling technology to overcome the above shortcomings.
Microdialysis is an in vivo sampling technique based on dialysis principles10. Under non-equilibrium conditions, the compounds to be measured are diffused and perfused from the tissue along the concentration gradient into the microdialysis probe embedded in the tissue into the dialysate, which is continuously removed along with the dialysate, achieving the purpose of sampling from the living tissue11,12. Compared with traditional sampling methods, the microdialysis technique has splendid advantages in the following aspects13,14,15: continuous real-time tracking of the changes of various compounds in blood; sampling requires no tedious pre-processing and can truly represent the concentration of the target compound at the sampling site; probes can be implanted into different parts of the body to investigate the absorb, distribution, metabolism, excretion and toxicity of the target compounds; the acquired sample contains no biological macromolecules (&#62;20 kD). Therefore, the higher quality blood samples ensure a better interpretation of CVDs and the mechanism treated by ethnic medicine.
Microdialysis sampling systems generally consist of micro-injection pumps, connecting tubes, animal-free movement tanks, microdialysis probes, and sample collectors16. As the most critical part of the device of the microdialysis system, common microdialysis probes comprise concentric probes, flexible probes, linear probes, and shunt probe17. Among these, flexible probes are soft and non-metallic probes, mainly used to collect samples from blood vessels and peripheral tissues such as heart, muscle, skin, and fat of awake and freely moving or anesthetized animals13. When in contact with blood vessels or tissues, the probe can be flexibly bent, thereby avoiding irreversible damage to the probe or sampling site. With the continuous development of probe technology, the application of microdialysis technology in various fields is also deepening. In this paper, the rat heart blood was dynamically and continuously acquired by the noninvasive microdialysis technology through the flexible probe designed for blood collection.

<table><tbody><tr><td>Animal anesthesia system</td><td>Rayward Life Technology Co., Ltd</td><td>R500IE</td><td></td></tr><tr><td>Animal temperature maintainer</td><td>Rayward Life Technology Co., Ltd</td><td>69020</td><td></td></tr><tr><td>Blood microdialysis probe</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>T55347</td><td></td></tr><tr><td>Catheter</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>T55347</td><td></td></tr><tr><td>Citrate</td><td>Merck Chemical Technology (Shanghai) Co., Ltd</td><td>251275</td><td></td></tr><tr><td>Electric shaver</td><td>Rayward Life Technology Co., Ltd</td><td>CP-5200</td><td></td></tr><tr><td>Fep tubing</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>3409501</td><td></td></tr><tr><td>Free movement tank for animals</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>CMA120</td><td></td></tr><tr><td>Glucose</td><td>Merck Chemical Technology (Shanghai) Co., Ltd</td><td>G8270</td><td></td></tr><tr><td>Hemostatic forceps</td><td>Rayward Life Technology Co., Ltd</td><td>F21020-16</td><td></td></tr><tr><td>Isofluran</td><td>Rayward Life Technology Co., Ltd</td><td>R510-22</td><td></td></tr><tr><td>Micro scissors</td><td>Beyotime Biotechnology Co., Ltd</td><td>FS221</td><td></td></tr><tr><td>Microdialysis collection tube</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>7431100</td><td></td></tr><tr><td>Microdialysis collector</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>CMA4004</td><td></td></tr><tr><td>Microdialysis in vitro stand</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>CMA130</td><td></td></tr><tr><td>Microdialysis microinjection pump</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>788130</td><td></td></tr><tr><td>Microdialysis syringe (1.0 mL)</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>8309020</td><td></td></tr><tr><td>Microdialysis tubing adapter</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>3409500</td><td></td></tr><tr><td>Microporous filter membrane</td><td>Merck Millipore Ltd.</td><td>R0DB36622</td><td></td></tr><tr><td>Non-absorbable surgical sutures</td><td>Shanghai Tianqing Biological Materials Co., Ltd</td><td>S19004</td><td></td></tr><tr><td>Operating table</td><td>Yuyan Scientific Instrument Co., Ltd</td><td>30153</td><td></td></tr><tr><td>Ophthalmic forceps</td><td>Rayward Life Technology Co., Ltd</td><td>F12016-15</td><td></td></tr><tr><td>Sodium citrate</td><td>Merck Chemical Technology (Shanghai) Co., Ltd</td><td>1613859</td><td></td></tr><tr><td>Sprague Dawley&amp;nbsp; (SD) rats</td><td>Chengdu Dossy Experimental Animals Co., Ltd</td><td>SYXK(&lt;img alt=&quot;Equation 1&quot; src=&quot;/files/ftp_upload/64531/64531eq01.jpg&quot; style=&quot;margin: auto;&quot;/&gt;)2019-049</td><td></td></tr><tr><td>Surgical scissors</td><td>Rayward Life Technology Co., Ltd</td><td>S14014-15</td><td></td></tr><tr><td>Surgical scissors</td><td>Shanghai Bingyu Fluid technology Co., Ltd</td><td>BY-103</td><td></td></tr><tr><td>Syringe needle</td><td>&amp;nbsp;CMA Microdialysis AB</td><td>T55347</td><td></td></tr><tr><td>Ultrasonic cleaner</td><td>Guangdong Goote Ultrasonic Co., Ltd</td><td>KMH1-240W8101</td><td></td></tr></tbody></table>

dynamic continuous blood extraction from rat heart via noninvasive microdialysis technique

Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique

Dynamic analysis of blood components is of great importance in understanding cardiovascular diseases and their related diseases, such as myocardial infarction, arrhythmia, atherosclerosis, cardiogenic pulmonary edema, pulmonary embolism, and cerebral embolism. At the same time, it is urgent to break through the continuous heart blood sampling technique in live rats to evaluate the effectiveness of distinctive ethnic medicine therapy. In this study, a blood microdialysis probe was implanted in the right jugular vein of rats in a precise and noninvasive surgical procedure. Cardiac blood samples were then collected at a rate of 2.87 nL/min to 2.98 mL/min by connecting to an online microdialysis sample collection system. Even more momentously, the acquired blood samples can temporarily be stored in microdialysis containers at 4 &#176;C. The microdialysis-based online continuous blood collection program from rat heart has greatly guaranteed the quality of blood samples, advancing and invigorating the scientific rationality of the research on systemic cardiovascular diseases and evaluating ethnomedicine therapy from the perspective of hematology.

The present protocol allowed obtaining the cardiac blood from conscious rats according to sampling parameters set in the microdialysis equipment. Normal blood samples must be bright red, while animals with hypoxia, potential blood clots, or anemic disease may have dark purple or dark red. Samples obtained through the blood microdialysis technique are colorless, clear, and transparent, which can be used to analyze the serum markers of different diseases and the blood distribution of drugs and their metabolites by employin...

The present protocol describes a simple and efficient method for the real-time and dynamic collection of rat heart blood using the microdialysis technique.

Dynamic analysis of blood components is of great importance in understanding cardiovascular diseases and their related diseases, such as myocardial ...

dynamic-continuous-blood-extraction-from-rat-heart-via-noninvasive

Research

JoVE Journal

Medicine

This protocol describes a method to collect cardiac interstitial fluid from the isolated, perfused rat heart. To physically separate interstitial transudate from coronary venous effluent perfusate, the Langendorff perfused heart is inverted, and the transudate (interstitial fluid) formed on the cardiac surface is collected using a soft latex cap.

The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

This article presents an efficient method to perform myocardial ischemia and subsequent chronic reperfusion in rats using a minimally invasive approach. In addition, left ventricular hemodynamic function of rats is assessed by echocardiography and isolated working heart...

Semi-Minimal Invasive Method to Induce Myocardial Infarction in Rats and the Assessment of Cardiac Function by an Isolated Working Heart System

The gold standard in cardiology for cellular and molecular functional experiments are cardiomyocytes. This article describes adaptations to the non-Langendorff technique to isolate mouse cardiomyocytes.

A Simple and Effective Method to Consistently Isolate Mouse Cardiomyocytes

The present protocol describes aortic cannulation and retrograde perfusion of the ex-vivo neonatal murine heart.  A two-person strategy, using a dissecting microscope and a blunted small gauge needle, permits reliable cannulation. Quantification of longitudinal contractile tension is achieved using a force transducer connected to the apex of the left ventricle.

Modified Technique for the Use of Neonatal Murine Hearts in the Langendorff Preparation

The protocol here provides a detailed real-time dynamic sampling of extracellular fluid from the hippocampus of awake rats using a microdialysis system.

Real-Time Dynamic Collection of Hippocampal Extracellular Fluid from Conscious Rats Using a Microdialysis System

The protocol shows repeated cerebrospinal fluid and blood collections from epileptic rats performed in parallel with continuous video-electroencephalogram (EEG) monitoring. These are instrumental for exploring possible links between changes in various body fluid molecules and seizure activity.

Sampling Cerebrospinal Fluid and Blood from Lateral Tail Vein in Rats During EEG Recordings

This protocol describes a method for measuring left ventricular pressure and volume using the pressure-volume conductance technique. This method enables continuous real-time monitoring of the effects of drugs on the heart.

Real-Time Detection of Ferulic Acid Effects on Rat Left Ventricle Using Pressure-Volume Conductivity Catheter

This protocol outlines advanced material fabrication and ex vivo rat heart methods for optical and electrical bidirectional biointerfacing, enabling precise cardiac stimulation, recording, and infarction modeling for bioelectronics research.

Bidirectional Electrical and Optoelectronic Interfaces in Healthy and Ischemic Ex Vivo Rat Hearts

Learn a non-invasive microdialysis technique for dynamic continuous blood extraction from rat hearts. This method enables real-time cardiovascular biomarker analysis without sample pre-processing.

Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique

Explore More Videos