Name: Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio (Video) | JoVE Journal
Uploaded: 2011-09-30T12:00:00.000+00:00
Duration: 6 min 43 s
Description: Learn heart dissection in zebrafish at larval, juvenile, and adult stages using standardized protocols for Danio rerio. This method enables precise morphological analysis and cardiac development research.

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5 minutes

Thanks to Marie Birne, Areti Tsiola, Jaymie Estevez and Arelys Uribe. This work was supported in part by CUNY Research Foundation and Howard Hughes Medical Institute Grant Summer Program for Undergraduate Students (SPUR) and NIH RO3 41702-00-01. Research was also supported by Queens College and some of the experiments were done on equipment from the Core Facility for Imaging, Cellular and Molecular Biology at Queens College.

Zebrafish were raised using standard protocols13 in the Queens College Animal Facility at 28°C. Individual fish pairs or group matings should be preformed and the resulting embryos cleaned and stored in a 28°C incubator. Fish health should be examined daily. At 5-day post fertilization (dpf), fish should be separated into tanks at a 10-15 fish density and entered into the fish facility nursery. For the protocol presented here zebrafish were collected at larva through adult stages: 15, 30, 45, 60, 90, 180 dpf. 
1. Zebrafish subject collection and fixation
<ol>
<li>Remove fish from habitat tank using net and place in 0.2% Tricaine to anesthetize.</li>
<li>Place anesthetized fish in ice water for 15 minutes to euthanize.</li>
<li>Make fresh 4% paraformaldehyde (PFA) in 1x Phosphate-buffered saline (PBS).</li>
<li>Incubate collected fish in 4% PFA for 1-2 hours at RT then overnight at 4°C in a petri dish wrapped in parafilm. This keeps the fish flat and makes later dissection much easier.</li>
<li>After fixation, rinse twice in PBS with Tween (PBT). Fish can be keep in PBT at 4°C for up to 10 days.</li>
</ol>
2. Measuring fish and dissection
<ol>
<li>Place single fish into petri dish half filled with fresh PBS and lay it on its right side.</li>
<li>Measure length of fish from snout to base of tail (the caudal peduncle), not including tail fins, using a digital mm caliper (Figure 1). This is the standard length (SL).</li>
<li>Each fish can be assigned a number/letter combination to allow an individual fish and the resulting dissected heart to be tracked for later analysis. Data, including parental stock number, age at fixation, SL measurements, were entered into a spreadsheet and organized based on these labels.</li>
<li>Before dissection, label PCR tube strips with assigned tracking number and fill with PBT or other solution appropriate for further analysis.</li>
<li>Orient the fish ventral side up in petri dish while stabilizing the body with forceps holding the head between the eyes and gills (Figure 2).</li>
</ol>
Smaller fish (smaller than 12mm SL):
<ol start="4">
<li>a) Using sharp forceps, or a micro-needle in a pin holder, remove the pectoral muscles and fins from the body to reveal the heart. The constant motion of the forceps during dissection can cause currents in the PBT that move the fish. This is particularly problematic for smaller fish, thus a micro-needle can be used to remove pectoral muscles and fins and open the body cavity.</li>
<li>a) Gently use the forceps to scoop the heart out of the cavity from under the atrium. Alternately, in some fish you may be able to remove the heart by gently pulling the bulbous out and the rest of the heart will follow. Be careful of the location of the atrium if using this technique as it can easily be damaged. If there is extra tissue that comes with the heart, that is fine and can be cleared after.</li>
<li>a) If the fish contains a fluorescent cardiac marker such as Tg(myl7:GFP)twu34, use fluorescent scope for dissection and verify removal of the heart by florescence (Figure 3).</li>
<li>a) Once the heart is removed from the cavity, use the forceps to hold the heart and a micro-needle to remove extra non-cardiac tissue and the epicardial lining from the outside of the heart.</li>
</ol>
Larger fish (greater than 12mm SL):
<ol start="4">
<li>b) Using spring handled micro-scissors, make three incisions of less than 1mm deep: 1- transverse cut through the gills 2- transverse cut at anterior belly 3- sagittal cut on ventral side connecting both cuts (Figure 4).</li>
<li>b) Using sharp forceps remove the pectoral muscles and fins from the body to open the body cavity and reveal the silvery tissue of the pericardium (Figure 5A). Remove this pericardial tissue and the heart will become visible (Figure 5B).</li>
<li>b) Use the micro-scissors to cut the artery connected to the bulbous arteriosus, located superior to the heart. Once this artery is cut, put the forceps tips under the atrium, use the forceps to scoop the heart out of the cavity. If there is extra tissue that comes with the heart, that is fine and can be removed later.</li>
<li>b) Once the heart is removed from the cavity, use the two forceps to hold the heart and remove extra tissue, or one pair of forceps to hold the heart and a micro-needle to remove extra, non-cardiac tissue. The epicardial lining, evident by its black pigmentation, can be removed from the outside of the heart if desired by gently scraping the tissue.</li>
</ol>
3. Photography of hearts
<ol>
<li>Prepare an 23g/L nutrient agar petri dish and cover solidified agar with PBS.</li>
<li>Using the forceps, make a well in the agar that allows the heart to rest in the desired orientation for photographing.</li>
<li>Remove the heart from the PCR tube by holding the tip of the bulbous with the forceps firmly. Place the heart in the agar dish.</li>
<li>Orient the heart in agar to photograph. The sample can be rotated after each picture to all possible orientations (Figure 6).</li>
<li>Use overhead light and short exposure time to photograph the hearts. Make adjustments for light quality using the exposure as necessary to obtain the best photograph.</li>
</ol>
4. Representative Results: 
An example of a well-dissected clean heart is shown in multiple orientations in Figure 6. Each heart will have some differences in overall morphology, but the heart should be complete, containing an intact ventricle, atrium and bulbous arteriosus. Some examples of this variation in overall morphology are shown in Figure 7. The black spotted epicardial tissue can also removed from the heart.
<img src="/files/ftp_upload/3165/3165fig1.jpg" alt="Zebrafish measurement using digital caliper, biology research, length analysis experiment." data-altupdatedat="1788235205000" data-alt="Figure 1"> Figure 1. Measure the fish from snout to caudal peduncle using a digital mm caliper. 
<img src="/files/ftp_upload/3165/3165fig2.jpg" alt="Catfish anatomy microscopic image; observe skin texture and fin structure for biological study." data-altupdatedat="1788235205000" data-alt="Figure 2"> Figure 2. Hold the fish with forceps between the eyes and gills.
<img src="/files/ftp_upload/3165/3165fig3.jpg" alt="Fluorescent microscopy image of transgenic tissue expressing GFP, showing cellular structures." data-altupdatedat="1788235205000" data-alt="Figure 3"> Figure 3. A cardiac specific fluorescent transgenic marker such as Tg(myl7:GFP)twu34, shown here, can aid in visualizing hearts dissected from smaller fish. 
<img src="/files/ftp_upload/3165/3165fig4.jpg" alt="Fish tissue microscopy image; includes sectioning lines for anatomical analysis and study." data-altupdatedat="1788235205000" data-alt="Figure 4"> Figure 4. Three incisions to remove the pectoral muscles and skin: 1- transverse cut through the gills 2- transverse cut at anterior belly 3- sagittal cut on ventral side connecting both cuts. 
<img src="/files/ftp_upload/3165/3165fig5.jpg" alt="Fish dissection showing internal anatomy in two stages; labeled arteries and veins in image B." data-altupdatedat="1788235205000" data-alt="Figure 5"> Figure 5. Black letters label the chambers of the heart: bulbous arteriosus (ba), ventricle (v), and atrium (a). 
A-	The upper body cavity is open to show the silvery tissue of the pericardium. 
B-	After removing the pericardium, the heart is visible and easily removed. 

<img src="/files/ftp_upload/3165/3165fig6.jpg" alt="	Embryo heart development stages, labeled images, anterior-posterior axis, anatomical orientation." data-altupdatedat="1788235205000" data-alt="Figure 6"> Figure 6. Different orientations of the heart are shown. Black letters label the chambers of the heart: bulbous arteriosus (ba), ventricle (v), and atrium (a). Grey arrows indicate location and orientation of the heart in reference to its normal location in the body: anterior (A), posterior (P), dorsal (D), ventral (V), left (L), and right (R). Scale bar indicates 400μm. 
A-	ventral view of the heart 
B-	dorsal view of the heart 
C-	lateral view from the left side 
D-	lateral view from the right side 

<img src="/files/ftp_upload/3165/3165fig7.jpg" alt="Embryo growth stages; six microscope images; developmental biology; &lt;12mm and &gt;12mm comparison." data-altupdatedat="1788235205000" data-alt="Figure 7"> Figure 7. Morphology of hearts of different sizes is shown. Black letters label the chambers of the heart: bulbous arteriosus (ba), ventricle (v), and atrium (a). Scale bar indicated 400 μm. 
	A-C- Hearts form fish with SL less than 12mm 
	D-F- Hearts form fish with SL greater than 12mm

<ol><li>Ahmad, N., Long, S., Rebagliati, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+southpaw+joins+the+roster%3A+the+role+of+the+zebrafish+nodal-related+gene+southpaw+in+cardiac+LR+asymmetry%5BTitle%5D)+AND+%22Trends+Cardiovasc+Med%22%5BJournal%5D)">A southpaw joins the roster: the role of the zebrafish nodal-related gene southpaw in cardiac LR asymmetry.</a> Trends Cardiovasc Med. 14, 43-49 (2004).</li>
<li>Armstrong, E. J., Bischoff, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Heart+valve+development%3A+endothelial+cell+signaling+and+differentiation%5BTitle%5D)+AND+%22Circ+Res%22%5BJournal%5D)">Heart valve development: endothelial cell signaling and differentiation.</a> Circ Res. 95, 459-470 (2004).</li>
<li>Bakkers, J., Verhoeven, M. C., Abdelilah-Seyfried, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Shaping+the+zebrafish+heart%3A+from+left-right+axis+specification+to+epithelial+tissue+morphogenesis%5BTitle%5D)+AND+%22Dev+Biol%22%5BJournal%5D)">Shaping the zebrafish heart: from left-right axis specification to epithelial tissue morphogenesis.</a> Dev Biol. 330, 213-220 (2009).</li>
<li>Glickman, N. S., Yelon, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cardiac+development+in+zebrafish%3A+coordination+of+form+and+function%5BTitle%5D)+AND+%22Semin+Cell+Dev+Biol%22%5BJournal%5D)">Cardiac development in zebrafish: coordination of form and function.</a> Semin Cell Dev Biol. 13, 507-513 (2002).</li>
<li>Heicklen-Klein, A., McReynolds, L. J., Evans, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Using+the+zebrafish+model+to+study+GATA+transcription+factors%5BTitle%5D)+AND+%22Semin+Cell+Dev+Biol%22%5BJournal%5D)">Using the zebrafish model to study GATA transcription factors.</a> Semin Cell Dev Biol. 16, 95-106 (2005).</li>
<li>Schoenebeck, J. J., Yelon, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Illuminating+cardiac+development%3A+Advances+in+imaging+add+new+dimensions+to+the+utility+of+zebrafish+genetics%5BTitle%5D)+AND+%22Semin+Cell+Dev+Biol%22%5BJournal%5D)">Illuminating cardiac development: Advances in imaging add new dimensions to the utility of zebrafish genetics.</a> Semin Cell Dev Biol. 18, 27-35 (2007).</li>
<li>Beis, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+and+cellular+analyses+of+zebrafish+atrioventricular+cushion+and+valve+development%5BTitle%5D)+AND+%22Development%22%5BJournal%5D)">Genetic and cellular analyses of zebrafish atrioventricular cushion and valve development.</a> Development. 132, 4193-4204 (2005).</li>
<li>Lepilina, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+dynamic+epicardial+injury+response+supports+progenitor+cell+activity+during+zebrafish+heart+regeneration%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">A dynamic epicardial injury response supports progenitor cell activity during zebrafish heart regeneration.</a> Cell. 127, 607-619 (2006).</li>
<li>Wills, A. A., Holdway, J. E., Major, R. J., Poss, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Regulated+addition+of+new+myocardial+and+epicardial+cells+fosters+homeostatic+cardiac+growth+and+maintenance+in+adult+zebrafish%5BTitle%5D)+AND+%22Development%22%5BJournal%5D)">Regulated addition of new myocardial and epicardial cells fosters homeostatic cardiac growth and maintenance in adult zebrafish.</a> Development. 135, 183-192 (2008).</li>
<li>Parichy, D. M., Elizondo, M. R., Mills, M. G., Gordon, T. N., Engeszer, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Normal+table+of+postembryonic+zebrafish+development%3A+staging+by+externally+visible+anatomy+of+the+living+fish%5BTitle%5D)+AND+%22Dev+Dyn%22%5BJournal%5D)">Normal table of postembryonic zebrafish development: staging by externally visible anatomy of the living fish.</a> Dev Dyn. 238, 2975-3015 (2009).</li>
<li>Huang, C. J., Tu, C. T., Hsiao, C. D., Hsieh, F. J., Tsai, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Germ-line+transmission+of+a+myocardium-specific+GFP+transgene+reveals+critical+regulatory+elements+in+the+cardiac+myosin+light+chain+2+promoter+of+zebrafish%5BTitle%5D)+AND+%22Dev+Dyn%22%5BJournal%5D)">Germ-line transmission of a myocardium-specific GFP transgene reveals critical regulatory elements in the cardiac myosin light chain 2 promoter of zebrafish.</a> Dev Dyn. 228, 30-40 (2003).</li>
<li>Westerfield, M. The Zebrafish Book, A guide for the laboratory use of zebrafish (Danio rerio). , University of Oregon Press. (1995).</li>
</ol>

No conflicts of interest declared.

These methods for dissection of the zebrafish heart of post-embryonic fish allows for the complete removal of an undamaged heart. While this technique is straightforward and provides guides for external cuts that will prevent damaging the heart, a steady hand is essential for smaller fish.
A key step during the dissection is to identify the silvery pericardium as the heart is directly within this sac. Visualizing the pericardium provides an important marker for finding and ultimately removing ...

<table><tbody><tr><td>Paraformaldehyde </td><td>EMD Millipore</td><td>PX0055</td><td>.04g/mL in PBS</td></tr><tr><td>Forceps #55 </td><td>Fine Science Tools</td><td>11295-51</td><td></td></tr><tr><td>Micro-scissors </td><td>Fine Science Tools</td><td>15005-08</td><td></td></tr><tr><td>Pin holder </td><td>Fine Science Tools</td><td>26016-12</td><td></td></tr><tr><td>Micro-needles </td><td>Fine Science Tools</td><td>10130-10</td><td></td></tr><tr><td>Nutrient Agar </td><td>BBL</td><td>11472</td><td>23g/L in distilled H&lt;sub&gt;2&lt;/sub&gt;O</td></tr><tr><td>Petri dishes</td><td>BD Biosciences</td><td>351029</td><td></td></tr><tr><td>PCR 8-tube strips </td><td>USA Scientific, Inc.</td><td>1402-2500</td><td></td></tr><tr><td>Digital Caliper</td><td>Fisher Scientific</td><td>14-648-17</td><td></td></tr></tbody></table>

heart dissection in larval, juvenile and adult zebrafish, danio rerio

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work examining post-embryonic through adult cardiac development has been limited7-10. Examining the changing morphology of the maturing and aging heart are restricted by the lack of techniques available for staging and isolating juvenile and adult hearts. In order to analyze heart development over the fish's lifespan, we dissect zebrafish hearts at numerous stages and photograph them for further analysis11. The morphological features of the heart can easily be quantified and individual hearts can be further analyzed by a host of standard methods. Zebrafish grow at variable rates and maturation correlates better with fish size than age, thus, post-fixation, we photograph and measure fish length as a gauge of fish maturation. This protocol explains two distinct, size dependent dissection techniques for zebrafish, ranging from larvae 3.5mm standard length (SL) with hearts of 100&mu;m ventricle length (VL), to adults, with SL of 30mm and VL 1mm or larger. Larval and adult fish have quite distinct body and organ morphology. Larvae are not only significantly smaller, they have less pigment and each organ is visually very difficult to identify. For this reason, we use distinct dissection techniques.
We used pre-dissection fixation procedures, as we discovered that hearts dissected directly after euthanization have a more variable morphology, with very loose and balloon like atria compared with hearts removed following fixation. The fish fixed prior to dissection, retain in vivo morphology and chamber position (data not shown). In addition, for demonstration purposes, we take advantage of the heart (myocardial) specific GFP transgenic Tg(myl7:GFP)twu34 (12), which allows us to visualize the entire heart and is particularly useful at early stages in development when the cardiac morphology is less distinct from surrounding tissues. Dissection of the heart makes further analysis of the cell and molecular biology underlying heart development and maturation using in situ hybridization, immunohistochemistry, RNA extraction or other analytical methods easier in post-embryonic zebrafish. This protocol will provide a valuable technique for the study of cardiac development maturation and aging.

A clear, standardized method for dissection and isolation of the zebrafish heart at multiple developmental stages are described. Annotation and quantification techniques are also discussed.

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work ...

The overall goal of this procedure is to remove the heart from the zebra fish. This is accomplished by first collecting and fixing zebra fish. Next, the body cavity is opened to visualize the heart.Then the heart is removed and cleaned of non-cardiac tissue. Finally, the heart is photographed. Ultimately, the morphology of the heart over time can be seen through microscopy, sectioning, and photography.Visual demonstration of this method is critical as the keyto dissection. Sta are difficult to learn because zebrafish are small and the tissues are not distinct in color and can be hard to identify To generate zebra fish from the larvae through adult stages. For these experiments, perform individual pear or group matings with adult fish.Clean the embryos and store them in a 28 degrees Celsius incubator. At five days post fertilization, separate fish into tanks at a 10 to 15 fish density and place them into the fish facility. Remove the fish from its tank at the appropriate age and place it in 0.2%Trica to anesthetize.Place the anesthetized fish in ice water for 15 minutes to euthanize. Meanwhile, add 4%para formaldehyde or PFA in one XPBS to a dish. Incubate collected fish in the 4%PFA for one to two hours at room temperature, then overnight at four degrees Celsius in a Petri dish wrapped in perfil.This keeps the fish flat and makes dissection much easier after fixation. Rinse twice in PBS with 0.1%Tween or PBT fish can be kept in PBT at four degrees Celsius for up to 10 days. Place a single fish into a Petri dish, half filled with fresh PBS and lay it on its right side.Measure the length of the fish from snout to the base of the tail, not including the tail fins using a digital millimeter caliper. This is the standard length or sl. Assign each fish a number letter combination to allow an individual fish and the resulting dissected heart to be tracked for later analysis.Enter all data, including all measurements into a spreadsheet and organized based on these labels before dissection. Label PCR tube strips with assigned tracking number and fill tubes with PVT or other solution appropriate for further analysis. Orient the fish ventral site up in a Petri dish while stabilizing the body with forceps, holding the head between the eyes and gills for fish shorter than 12 millimeters in standard length.Using sharp forceps or a microneedle in a pin holder. Remove the pectoral muscles and fins from the body to reveal the heart. The constant motion of the forceps during dissection can cause currents in the PPT that move the fish.This is particularly problematic for smaller fish. Thus, a microneedle needs to be used to remove pectoral muscles and fin and open the body cavity. Use the forceps to gently scoop the heart out of the cavity from under the atrium.If the fish contains a fluorescent cardiac marker, use a fluorescent scope for the dissection and verify removal of the heart by fluorescence. Once the heart is removed from the cavity, use the forceps to hold the heart and a microneedle to remove extra noncardiac tissue and the epicardial lining from the outside of the heart. For fish longer than 12 millimeters in standard length.Using spring handled micro scissors, make three incisions. First, make a transverse cut through the gills. Second, make another transverse cut at the anterior belly.And third, make a sagittal cut on the ventral side connecting both transverse cuts. Using sharp forceps, remove the pectoral muscles and fins from the body to open the body cavity and reveal the silvery tissue of the pericardium. Remove this pericardial tissue and the heart will become visible.Use the micro scissors to cut the artery connected to the bulbus arteriosis located superior to the heart. Once this artery is cut, place the tips of the forceps under the atrium and use the forceps to scoop the heart out of the cavity. If there is extra tissue that comes with the heart, that is fine as it can be removed later.Alternatively, in some fish, it is possible to remove the heart by gently pulling the bulbus out and the rest of the heart will follow. Be careful of the location of the atrium if using the alternative technique as it can easily be damaged. If there's extra tissue that comes with the heart, this is fine and can be cleared after.Once the heart is removed from the cavity, use the two forceps to hold the heart and remove the extra tissue or one pair of forceps to hold the heart and a microneedle to remove extra non-cardiac tissue. To photograph the zebrafish heart, prepare a 23 gram per liter nutrient agar petri dish and cover the solidified agar with PBS. Use the forceps to make a well in the agar that allows the heart to fit in the proper orientation.Remove the heart from the PCR tube by holding the tip of the bulbus. With the forceps firmly. Place the heart in the agar dish.Orient the heart in the agar to photograph. The sample can be rotated after each picture to all possible orientations. Use overhead, light, and short exposure times to photograph the hearts.Make adjustments for light quality using the exposure as necessary to obtain the best photograph. An example of a well dissected clean heart is shown here. Each heart will have some differences in overall morphology, but the heart should be complete containing an intact ventricle atrium and bulbous arteriosis after its development.This technique paved the way for researchers in the field of zebrafish cardiac development to explore cardiac maturation in various stages of zebrafish development.

heart-dissection-in-larval-juvenile-and-adult-zebrafish-danio-rerio

Research

JoVE Journal

Biology

A rapid way to conduct immunostaining of zebrafish embryonic heart is described.  Compared to the whole mount immunostaining approach, this method dramatically increases the penetration of the antibodies, which allows obtaining high resolution images that reveal cellular/subcellular structures in the heart within a much reduced processing time.

Immunostaining of Dissected Zebrafish Embryonic Heart 

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Heart Dissection: A Method to Observe Zebrafish Cardiac Development

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Large-scale Zebrafish Embryonic Heart Dissection for Transcriptional Analysis

Use of zebrafish for cardiovascular research is expanding towards research on adult hearts. For these applications, quick and simple isolation of cardiac tissues is key to avoid post-mortem changes and to obtain an adequate number of samples. Here, we describe a fast and reproducible method for dissecting adult zebrafish hearts.

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Here, we present a reliable, minimally invasive, and cost-effective method to record and interpret electrocardiograms in live anesthetized adult zebrafish.

In Vivo Surface Electrocardiography for Adult Zebrafish

The goal of this protocol is to detail a novel method for the assessment of segmental cardiac function in embryonic zebrafish under both physiological and pathological conditions.

Displacement Analysis of Myocardial Mechanical Deformation (DIAMOND) Reveals Segmental Heterogeneity of Cardiac Function in Embryonic Zebrafish

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High-Frequency Ultrasound Echocardiography to Assess Zebrafish Cardiac Function

We describe optimized tools to study the zebrafish heart in vivo with light sheet fluorescence microscopy. Specifically, we suggest bright cardiac transgenic lines and present new gentle embedding and immobilization techniques that avoid developmental and heart defects. A possible data acquisition and analysis pipeline adapted to cardiac imaging is also provided.

Light Sheet Microscopy of Fast Cardiac Dynamics in Zebrafish Embryos

The present protocol describes the acute isolation of viable cardiac and vascular smooth muscle cells from adult, juvenile, larval, and embryonic zebrafish (Danio rerio), suitable for electrophysiological studies.

Isolation of Cardiac and Vascular Smooth Muscle Cells from Adult, Juvenile, Larval and Embryonic Zebrafish for Electrophysiological Studies

This protocol describes a low-cost light microscopy method to assess cardiac morphology and function in zebrafish embryos, enabling reproducible evaluation of developmental cardiotoxicity without the need for advanced imaging systems.

Functional Cardiac Imaging in Zebrafish Embryos Using Standard Microscopy and Video Analysis: Applications in Environmental and Biomedical Research

This study establishes a sensitive immunofluorescence-based imaging method to quantify global DNA methylation in individual zebrafish embryonic hearts. This approach overcomes the challenge of limited tissue material and bypasses the need for bisulfite conversion.

Detection of Global DNA Methylation in the Hearts of Zebrafish Larvae

Learn heart dissection in zebrafish at larval, juvenile, and adult stages using standardized protocols for Danio rerio. This method enables precise morphological analysis and cardiac development research.

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

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