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Optical transfection
open-in-new
<h2 id="terminology">Terminology</h2> <p>The meaning of the term <a href="/facts/Transfection/R94fLSQF">transfection</a> has evolved.<a class="footnote-ref" id="fnref:16" href="#fn:16"><sup>16</sup></a> The original meaning of transfection was "infection by transformation", <i>i.e.</i> introduction of DNA (or RNA) from a prokaryote-infecting virus or <a href="/facts/Bacteriophage/L2Ll0T3q">bacteriophage</a> into cells, resulting in an infection. Because the term transformation had another sense in animal cell biology (a genetic change allowing long-term propagation in culture, or acquisition of properties typical of cancer cells), the term transfection acquired, for animal cells, its present meaning of a change in cell properties caused by introduction of <a href="/facts/DNA/nB89Iauz">DNA</a> (or other nucleic acid species such as <a href="/facts/RNA/HxPeNfNj">RNA</a> or <a href="/facts/SiRNA/891ytkD9">SiRNA</a>). </p><p>Because of this strict definition of <a href="/facts/Transfection/R94fLSQF">transfection</a>, optical transfection also refers only to the introduction of nucleic acid species. The introduction of other impermeable compounds into a cell, such as organic <a href="/facts/Fluorophore/wDWBmYhn">fluorophores</a> or semiconductor <a href="/facts/Quantum_dot/K5Ytv8Yw">quantum nanodots</a> is not strictly speaking "transfection," and is therefore referred to as "optical injection" or one of the many other terms now outlined. </p><p>The lack of a unified name for this technology makes reviewing the literature on the subject very difficult.<a class="footnote-ref" id="fnref:17" href="#fn:17"><sup>17</sup></a> Optical injection has been described using over a dozen different names or phrases (see bulleted lists below). Some trends in the literature are clear. The first term of the technique is invariably a derivation of word laser, optical, or photo, and the second term is usually in reference to injection, transfection, poration, perforation or puncture. Like many cellular perturbations, when a single cell or group of cells is treated with a laser, three things can happen: the cell dies (overdose), the cell membrane is permeabilised, substances enter, and the cell recovers (therapeutic dose), or nothing happens (underdose). There have been suggestions in the literature to reserve the term optoinjection for when a therapeutic dose is delivered upon a single cell,<a class="footnote-ref" id="fnref:18" href="#fn:18"><sup>18</sup></a><a class="footnote-ref" id="fnref:19" href="#fn:19"><sup>19</sup></a><a class="footnote-ref" id="fnref:20" href="#fn:20"><sup>20</sup></a> and the term optoporation for when a laser generated shockwave treats a cluster of many (10s to 100s) cells.<a class="footnote-ref" id="fnref:21" href="#fn:21"><sup>21</sup></a><a class="footnote-ref" id="fnref:22" href="#fn:22"><sup>22</sup></a><a class="footnote-ref" id="fnref:23" href="#fn:23"><sup>23</sup></a><a class="footnote-ref" id="fnref:24" href="#fn:24"><sup>24</sup></a> The first definition of optoinjection is uncontroversial. The definition of optoporation, however, has failed to be adopted, with a similar number of references using the term to denote the dosing of single cells<a class="footnote-ref" id="fnref:25" href="#fn:25"><sup>25</sup></a><a class="footnote-ref" id="fnref:26" href="#fn:26"><sup>26</sup></a><a class="footnote-ref" id="fnref:27" href="#fn:27"><sup>27</sup></a><a class="footnote-ref" id="fnref:28" href="#fn:28"><sup>28</sup></a> as those using the term to denote the simultaneous dosing of clusters of many cells <a class="footnote-ref" id="fnref:29" href="#fn:29"><sup>29</sup></a><a class="footnote-ref" id="fnref:30" href="#fn:30"><sup>30</sup></a><a class="footnote-ref" id="fnref:31" href="#fn:31"><sup>31</sup></a><a class="footnote-ref" id="fnref:32" href="#fn:32"><sup>32</sup></a> </p><p>As the field stands, it is the opinion of the authors of a review article on the subject<a class="footnote-ref" id="fnref:33" href="#fn:33"><sup>33</sup></a> that the term optoinjection always be included as a keyword in future publications, regardless of their own naming preferences. </p><p>Terms agreed by consensus </p> <ul><li>Optoinjection (or any derivations of laser injection, optical injection, photoinjection): The transfer of any membrane impermeable substance into a cell using light. A general term that also encompasses optical transfection.</li> <li>Optical transfection (or any derivations of laser transfection, optotransfection, phototransfection): A specific type of optical transfection - the transfer of nucleic acids into a cell using light for the purposes of eliciting protein translation from those acids. To be in line with the current definition of transfection in the biological community, non-nucleic acids (such as fluorophores) cannot, by definition, be optically transfected (only optically injected).</li> <li>Photoporation (or any derivations of [laser-] or [optical-] or [opto-] or [photo-] AND [ poration] or [-permeabilisation] or [-puncture] or [-perforation]): The generation of a transient hole or holes on the plasma membrane (or cell wall) of a cell usually for the purpose of optical injection. See possible exception: Optoporation</li> <li>-surgery (such as cell nanosurgery, laser nanosurgery, laser surgery): A general term that incorporates all of the above definitions, but also includes the concepts of the ablation or optical manipulation of cell material for other purposes besides pore generation. Examples include selective cell ablation to purify cell populations, chromosome dissection, cytoskeleton disruption, organelle ablation, axotomy,<a class="footnote-ref" id="fnref:34" href="#fn:34"><sup>34</sup></a> or the optical tweezing or isolation of intracellular material.</li></ul> <p>Terms under deliberation </p> <ul><li>Optoporation: Has been suggested to mean the dosing of a cluster of cells with a shockwave mediated mechanism, which usually results in a doughnut shaped therapeutic zone.<a class="footnote-ref" id="fnref:35" href="#fn:35"><sup>35</sup></a><a class="footnote-ref" id="fnref:36" href="#fn:36"><sup>36</sup></a><a class="footnote-ref" id="fnref:37" href="#fn:37"><sup>37</sup></a><a class="footnote-ref" id="fnref:38" href="#fn:38"><sup>38</sup></a> On the contrary, has also been synonymously used with the term photoporation.<a class="footnote-ref" id="fnref:39" href="#fn:39"><sup>39</sup></a><a class="footnote-ref" id="fnref:40" href="#fn:40"><sup>40</sup></a><a class="footnote-ref" id="fnref:41" href="#fn:41"><sup>41</sup></a><a class="footnote-ref" id="fnref:42" href="#fn:42"><sup>42</sup></a></li> <li>Laserfection: Has been suggested to mean the dosing of a cluster of cells with a circularly shaped therapeutic zone. Term reserved for Cyntellect's laser-enabled analysis and processing (LEAP) system.</li> <li>Light-induced convective transmembrane transport: A newly coined term for optoinjection.<a class="footnote-ref" id="fnref:43" href="#fn:43"><sup>43</sup></a></li></ul> <p>Some of the above was reproduced with permission from.<a class="footnote-ref" id="fnref:44" href="#fn:44"><sup>44</sup></a> </p> <h2 id="methods">Methods</h2> <p>A typical optical transfection protocol is as follows:<a class="footnote-ref" id="fnref:45" href="#fn:45"><sup>45</sup></a> 1) Build an <a href="/facts/Optical_tweezers/wsPxwmX6">optical tweezers</a> system with a high NA objective 2) Culture cells to 50-60% confluency 3) Expose cells to at least 10 μg/mL of plasmid DNA 4) Dose the plasma membrane of each cell with 10-40 ms of focussed laser, at a power of <100 mW at focus 5) Observe transient transfection 24-96h later 6) Add <a href="/facts/Growth_medium/97IMkpFP">selective medium</a> if the generation of stable colonies is desired </p> <h2 id="see-also">See also</h2> <ul><li><a href="/facts/Transfection/R94fLSQF">Transfection</a></li> <li><a href="/facts/Transformation_(genetics)/n82cCRCu">Transformation</a></li> <li><a href="/facts/Transduction_(genetics)/nk0833JV">Transduction</a></li> <li><a href="/facts/Cationic_liposome/by0CD6uH">Cationic liposome</a></li> <li><a href="/facts/Nucleofection/tBwzXxSy">Nucleofection</a></li> <li><a href="/facts/Magnet_assisted_transfection/k9nLGtNk">Magnet assisted transfection</a></li></ul> <h2 id="external-links">External links</h2> <ul><li><a href="http://photon.st-andrews.ac.uk/biophotonics/index.php/research/optical-transfection">Research in optical transfection at the University of St Andrews</a></li> <li><a href="https://meshb.nlm.nih.gov/record/ui?name=Transfection">Transfection</a> at the U.S. National Library of Medicine <a href="/facts/Medical_Subject_Headings/C57g2JuF">Medical Subject Headings</a> (MeSH)</li> <li><a href="http://www.nature.com/nmeth/journal/v2/n11/full/nmeth1105-875.html">Overview of transfection methods</a> in <i>Nature Methods</i> 2, 875 - 883 (2005)</li></ul> <h2 id="references">References</h2> <ol> <li id="fn:1"><p>Tsukakoshi, M.; Kurata, S.; Nomiya, Y.; Ikawa, Y.; Kasuya, T. (1984). "A novel method of DNA transfection by laser microbeam cell surgery". Applied Physics B: Photophysics and Laser Chemistry. 35 (3). Springer Science and Business Media LLC: 135–140. Bibcode:1984ApPhB..35..135T. doi:10.1007/bf00697702. ISSN 0721-7269. S2CID 123250337. <a href="/wiki/Bibcode_(identifier)" target="_blank">/wiki/Bibcode_(identifier)</a> <a href="#fnref:1" class="footnote-back-ref">↩</a></p></li> <li id="fn:2"><p>Paterson, L.; Agate, B.; Comrie, M.; Ferguson, R.; Lake, T. K.; et al. (2005). "Photoporation and cell transfection using a violet diode laser". Optics Express. 13 (2). The Optical Society: 595–600. Bibcode:2005OExpr..13..595P. doi:10.1364/opex.13.000595. ISSN 1094-4087. PMID 19488389. <a href="https://doi.org/10.1364%2Fopex.13.000595" target="_blank">https://doi.org/10.1364%2Fopex.13.000595</a> <a href="#fnref:2" class="footnote-back-ref">↩</a></p></li> <li id="fn:3"><p>Palumbo, Giuseppe; Caruso, Matilde; Crescenzi, Elvira; Tecce, Mario F.; Roberti, Giuseppe; Colasanti, Alberto (1996). "Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation". Journal of Photochemistry and Photobiology B: Biology. 36 (1). Elsevier BV: 41–46. Bibcode:1996JPPB...36...41P. doi:10.1016/s1011-1344(96)07335-6. ISSN 1011-1344. PMID 8988610. <a href="/wiki/Bibcode_(identifier)" target="_blank">/wiki/Bibcode_(identifier)</a> <a href="#fnref:3" class="footnote-back-ref">↩</a></p></li> <li id="fn:4"><p>Tsampoula, X.; Taguchi, K.; Čižmár, T.; Garces-Chavez, V; Ma, N.; et al. (2008-10-10). "Fibre based cellular transfection". Optics Express. 16 (21). The Optical Society: 17007–13. Bibcode:2008OExpr..1617007T. doi:10.1364/oe.16.017007. ISSN 1094-4087. PMID 18852810. <a href="https://doi.org/10.1364%2Foe.16.017007" target="_blank">https://doi.org/10.1364%2Foe.16.017007</a> <a href="#fnref:4" class="footnote-back-ref">↩</a></p></li> <li id="fn:5"><p>Uchugonova, Aisada; König, Karsten; Bueckle, Rainer; Isemann, Andreas; Tempea, Gabriel (2008-06-11). "Targeted transfection of stem cells with sub-20 femtosecond laser pulses". Optics Express. 16 (13). The Optical Society: 9357–64. Bibcode:2008OExpr..16.9357U. doi:10.1364/oe.16.009357. ISSN 1094-4087. PMID 18575499. <a href="https://doi.org/10.1364%2Foe.16.009357" target="_blank">https://doi.org/10.1364%2Foe.16.009357</a> <a href="#fnref:5" class="footnote-back-ref">↩</a></p></li> <li id="fn:6"><p>Brown, C. T. A.; Stevenson, D. J.; Tsampoula, X.; McDougall, C.; Lagatsky, A. A.; et al. (2008). "Enhanced operation of femtosecond lasers and applications in cell transfection". Journal of Biophotonics. 1 (3). Wiley: 183–199. doi:10.1002/jbio.200810011. ISSN 1864-063X. PMID 19412968. S2CID 10801756. <a href="https://doi.org/10.1002%2Fjbio.200810011" target="_blank">https://doi.org/10.1002%2Fjbio.200810011</a> <a href="#fnref:6" class="footnote-back-ref">↩</a></p></li> <li id="fn:7"><p>Baumgart, J.; Bintig, W.; Ngezahayo, A.; Willenbrock, S.; Murua Escobar, H.; Ertmer, W.; Lubatschowski, H.; Heisterkamp, A. (2008). "Quantified femtosecond laser based opto-perforation of living GFSHR-17 and MTH53 a cells". Optics Express. 16 (5). The Optical Society: 3021–31. Bibcode:2008OExpr..16.3021B. doi:10.1364/oe.16.003021. ISSN 1094-4087. PMID 18542388. <a href="https://doi.org/10.1364%2Foe.16.003021" target="_blank">https://doi.org/10.1364%2Foe.16.003021</a> <a href="#fnref:7" class="footnote-back-ref">↩</a></p></li> <li id="fn:8"><p>Lei, Ming; Xu, Hanpeng; Yang, Hao; Yao, Baoli (2008). "Femtosecond laser-assisted microinjection into living neurons". Journal of Neuroscience Methods. 174 (2). Elsevier BV: 215–218. doi:10.1016/j.jneumeth.2008.07.006. ISSN 0165-0270. PMID 18687359. S2CID 10242427. <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:8" class="footnote-back-ref">↩</a></p></li> <li id="fn:9"><p>Tsampoula, X.; Garcés-Chávez, V.; Comrie, M.; Stevenson, D. J.; Agate, B.; Brown, C. T. A.; Gunn-Moore, F.; Dholakia, K. (2007-07-30). "Femtosecond cellular transfection using a nondiffracting light beam". Applied Physics Letters. 91 (5). AIP Publishing: 053902–053903. Bibcode:2007ApPhL..91e3902T. doi:10.1063/1.2766835. ISSN 0003-6951. <a href="/wiki/Bibcode_(identifier)" target="_blank">/wiki/Bibcode_(identifier)</a> <a href="#fnref:9" class="footnote-back-ref">↩</a></p></li> <li id="fn:10"><p>Peng, Cheng; Palazzo, Robert E.; Wilke, Ingrid (2007-04-03). "Laser intensity dependence of femtosecond near-infrared optoinjection". Physical Review E. 75 (4). American Physical Society (APS): 041903. Bibcode:2007PhRvE..75d1903P. doi:10.1103/physreve.75.041903. ISSN 1539-3755. PMID 17500917. <a href="/wiki/Bibcode_(identifier)" target="_blank">/wiki/Bibcode_(identifier)</a> <a href="#fnref:10" class="footnote-back-ref">↩</a></p></li> <li id="fn:11"><p>Stevenson, D.; Agate, B.; Tsampoula, X.; Fischer, P.; Brown, C. T. A.; Sibbett, W.; Riches, A.; Gunn-Moore, F.; Dholakia, K. (2006). "Femtosecond optical transfection of cells:viability and efficiency". Optics Express. 14 (16). The Optical Society: 7125–33. Bibcode:2006OExpr..14.7125S. doi:10.1364/oe.14.007125. ISSN 1094-4087. PMID 19529083. <a href="https://doi.org/10.1364%2Foe.14.007125" target="_blank">https://doi.org/10.1364%2Foe.14.007125</a> <a href="#fnref:11" class="footnote-back-ref">↩</a></p></li> <li id="fn:12"><p>Barrett, Lindy E; Sul, Jai-Yoon; Takano, Hajime; Van Bockstaele, Elisabeth J; Haydon, Philip G; Eberwine, James H (2006-05-23). "Region-directed phototransfection reveals the functional significance of a dendritically synthesized transcription factor". Nature Methods. 3 (6). Springer Science and Business Media LLC: 455–460. doi:10.1038/nmeth885. ISSN 1548-7091. PMID 16721379. S2CID 10536176. <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:12" class="footnote-back-ref">↩</a></p></li> <li id="fn:13"><p>Tirlapur, Uday K.; König, Karsten (2002). "Targeted transfection by femtosecond laser". Nature. 418 (6895). Springer Science and Business Media LLC: 290–291. doi:10.1038/418290a. ISSN 0028-0836. PMID 12124612. S2CID 4370674. <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:13" class="footnote-back-ref">↩</a></p></li> <li id="fn:14"><p>Soughayer, Joseph S.; Krasieva, Tatiana; Jacobson, Stephen C.; Ramsey, J. Michael; Tromberg, Bruce J.; Allbritton, Nancy L. (2000). "Characterization of Cellular Optoporation with Distance". Analytical Chemistry. 72 (6). American Chemical Society (ACS): 1342–1347. doi:10.1021/ac990982u. ISSN 0003-2700. PMID 10740880. S2CID 6373605. <a href="https://escholarship.org/uc/item/41m9w7s0" target="_blank">https://escholarship.org/uc/item/41m9w7s0</a> <a href="#fnref:14" class="footnote-back-ref">↩</a></p></li> <li id="fn:15"><p>Mohanty, Samarendra K.; Sharma, Mrinalini; Gupta, Pradeep K. (2003). "Laser-assisted microinjection into targeted animal cells". Biotechnology Letters. 25 (11). Springer Science and Business Media LLC: 895–899. doi:10.1023/a:1024038609045. ISSN 0141-5492. PMID 12889802. S2CID 33912519. <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:15" class="footnote-back-ref">↩</a></p></li> <li id="fn:16"><p>"Transfection" at Dorland's Medical Dictionary <a href="https://web.archive.org/web/20090628224336/http://www.mercksource.com/pp/us/cns/cns_hl_dorlands_split.jsp?pg=/ppdocs/us/common/dorlands/dorland/eight/000110186.htm" target="_blank">https://web.archive.org/web/20090628224336/http://www.mercksource.com/pp/us/cns/cns_hl_dorlands_split.jsp?pg=/ppdocs/us/common/dorlands/dorland/eight/000110186.htm</a> <a href="#fnref:16" class="footnote-back-ref">↩</a></p></li> <li id="fn:17"><p>Stevenson, David J.; Gunn-Moore, Frank J.; Campbell, Paul; Dholakia, Kishan (2010-01-11). "Single cell optical transfection". Journal of the Royal Society Interface. 7 (47). The Royal Society: 863–871. doi:10.1098/rsif.2009.0463. ISSN 1742-5689. PMC 2871806. PMID 20064901. <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871806" target="_blank">https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871806</a> <a href="#fnref:17" class="footnote-back-ref">↩</a></p></li> <li id="fn:18"><p>Krasieva, Tatiana B.; Chapman, Curtis F.; LaMorte, Vickie J.; Venugopalan, Vasan; Berns, Michael W.; Tromberg, Bruce J. (1998-04-29). "Cell permeabilization and molecular transport by laser microirradiation". In Farkas, Daniel L.; Leif, Robert C.; Tromberg, Bruce J. (eds.). Optical Investigations of Cells in Vitro and in Vivo. Vol. 3260. SPIE. pp. 38–44. doi:10.1117/12.307113. S2CID 59321515. {{cite book}}: |journal= ignored (help) <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:18" class="footnote-back-ref">↩</a></p></li> <li id="fn:19"><p>Rhodes, Kate; Clark, Imran; Zatcoff, Michelle; Eustaquio, Trisha; Hoyte, Kwame L.; Koller, Manfred R. (2007). "Cellular Laserfection". Methods in Cell Biology. Vol. 82. Elsevier. pp. 309–333. doi:10.1016/s0091-679x(06)82010-8. ISBN 978-0-12-370648-5. ISSN 0091-679X. PMID 17586262. <a href="978-0-12-370648-5" target="_blank">978-0-12-370648-5</a> <a href="#fnref:19" class="footnote-back-ref">↩</a></p></li> <li id="fn:20"><p>Venugopalan, Vasan; Guerra, Arnold; Nahen, Kester; Vogel, Alfred (2002-02-04). "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation". Physical Review Letters. 88 (7). American Physical Society (APS): 078103. Bibcode:2002PhRvL..88g8103V. doi:10.1103/physrevlett.88.078103. ISSN 0031-9007. PMID 11863944. S2CID 6235612. <a href="http://www.escholarship.org/uc/item/9309n2wn" target="_blank">http://www.escholarship.org/uc/item/9309n2wn</a> <a href="#fnref:20" class="footnote-back-ref">↩</a></p></li> <li id="fn:21"><p>Krasieva, Tatiana B.; Chapman, Curtis F.; LaMorte, Vickie J.; Venugopalan, Vasan; Berns, Michael W.; Tromberg, Bruce J. (1998-04-29). "Cell permeabilization and molecular transport by laser microirradiation". In Farkas, Daniel L.; Leif, Robert C.; Tromberg, Bruce J. (eds.). Optical Investigations of Cells in Vitro and in Vivo. Vol. 3260. SPIE. pp. 38–44. doi:10.1117/12.307113. S2CID 59321515. {{cite book}}: |journal= ignored (help) <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:21" class="footnote-back-ref">↩</a></p></li> <li id="fn:22"><p>Rhodes, Kate; Clark, Imran; Zatcoff, Michelle; Eustaquio, Trisha; Hoyte, Kwame L.; Koller, Manfred R. (2007). "Cellular Laserfection". Methods in Cell Biology. Vol. 82. Elsevier. pp. 309–333. doi:10.1016/s0091-679x(06)82010-8. ISBN 978-0-12-370648-5. ISSN 0091-679X. PMID 17586262. <a href="978-0-12-370648-5" target="_blank">978-0-12-370648-5</a> <a href="#fnref:22" class="footnote-back-ref">↩</a></p></li> <li id="fn:23"><p>Soughayer, Joseph S.; Krasieva, Tatiana; Jacobson, Stephen C.; Ramsey, J. Michael; Tromberg, Bruce J.; Allbritton, Nancy L. (2000). "Characterization of Cellular Optoporation with Distance". Analytical Chemistry. 72 (6). American Chemical Society (ACS): 1342–1347. doi:10.1021/ac990982u. ISSN 0003-2700. PMID 10740880. S2CID 6373605. <a href="https://escholarship.org/uc/item/41m9w7s0" target="_blank">https://escholarship.org/uc/item/41m9w7s0</a> <a href="#fnref:23" class="footnote-back-ref">↩</a></p></li> <li id="fn:24"><p>Venugopalan, Vasan; Guerra, Arnold; Nahen, Kester; Vogel, Alfred (2002-02-04). "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation". Physical Review Letters. 88 (7). American Physical Society (APS): 078103. Bibcode:2002PhRvL..88g8103V. doi:10.1103/physrevlett.88.078103. ISSN 0031-9007. PMID 11863944. 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