TY - JOUR AU - Parnell, Dakota James AU - Orell, Jesse Reed AU - Ali, Syed Monis AU - Walters, James William PY - 2020/04/29 Y2 - 2024/03/19 TI - Optimizing a micro-fluidic slide for in vivo imaging of larval Zebrafish. JF - Proceedings of the West Virginia Academy of Science JA - Proc. W. Va. Acad. Sci. VL - 92 IS - 1 SE - Meeting Abstracts-Poster DO - 10.55632/pwvas.v92i1.736 UR - https://pwvas.org/index.php/pwvas/article/view/736 SP - AB - <span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">Larval zebrafish (Danio </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">rerio</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">) are optically transparent at six- and seven-days post fertilization (</span><span class="SpellingError SCXW154402139 BCX0" data-ccp-parastyle="annotation text">dpf</span><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">) which makes them </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="none"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">useful research models for the study of metabolic and physiological processes. </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">Previous </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">methods to image </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">z</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">ebrafish </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">were</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> limited to embryogenesis </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">and early larval stages. </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">Our goal</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">was</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> to design a novel micro-fluidic</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> slide </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">for mounting zebrafish </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">that</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> greatly extend</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">s</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">the duration of time </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">for </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">in vivo </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">imaging of zebrafish </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">larvae </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">that are</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> six to seven </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">days</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> post fertilization (</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="SpellingError SCXW154402139 BCX0" data-ccp-parastyle="annotation text">dpf</span><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">). </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">The </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">ideal mounting slide will have channels to provide</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> media</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> exchange</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">, </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">nourishment, and</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">/or</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">drug</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> treatments</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> and </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">channels to remove waste and </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">flush the system</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> of </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">previous test conditions</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">.</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">We had a microfluidic slide manufactured to our specifications. Initial testing revealed that the larval bed was too large.</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">To optimize our previous prototype, we used the </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">Zeiss</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> Discovery V8 microscope to recalculate our measurements for the larvae bed. </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">We have now re-imaged five each of six </span><span class="SpellingError SCXW154402139 BCX0" data-ccp-parastyle="annotation text">dpf</span><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> larvae and five each of seven </span><span class="SpellingError SCXW154402139 BCX0" data-ccp-parastyle="annotation text">dpf</span><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> larvae in order to adjust the measurements to the new average larval </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">size</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="auto"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">. </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="none"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text">This work was</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="none"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> supported by NIH Grant P20GM103434 to the West Virginia </span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="none"><span class="SpellingError SCXW154402139 BCX0" data-ccp-parastyle="annotation text">IDeA</span></span><span class="TextRun SCXW154402139 BCX0" lang="EN-US" xml:lang="EN-US" data-contrast="none"><span class="NormalTextRun SCXW154402139 BCX0" data-ccp-parastyle="annotation text"> Network for Biomedical Research Excellence and NIH Grant P20GM103434 awarded to Bluefield State College</span></span> ER -