Index 577..592

A A23187 (Ca2þ ionophore), 215, 216 Acetamidoiminodiacetic acid (ADA), affinity for Ca2þ, 537t Acetoxymethyl (AM) esters of Ca2þ indicators, 408. See also specific indicators Acids disposal of, 573 safe handling of, 572, 575 Acridine orange, 521–523 ACSF for calcium imaging (recipe), 149, 152 Action potentials, back propagating, 142, 142f Activation of Ca2þ-dependent ion channels, 79–84 ADA (acetamidoiminodiacetic acid), affinity for Ca2þ, 537t Adenophostin A, 173–174, 173f, 174f Adenosine diphosporibose (ADPR), 498 ADP, affinity for Ca2þ, 537t ADP-ribosyl cyclase, 490, 493, 498–501, 518, 530 Aequorin, 177–210, 193, 465–466 advantages of, 180, 183, 194 AquaLite, 203 calibration curve, 179, 187, 188f as Ca2þ-sensitive photoprotein, 178 chimeras, 181t–182t decline in use as cytosolic probe, 179 disadvantages of, 183 experimental applications, 179–183 f-aequorin, 203 hcp-aequorin, 203 history of, 177–178 introduction into zebrafish embryos, 193–210, 195f monitoring yeast intracellular Ca2þ levels using in vivo bioluminescence assay (protocol), 456–459 materials, 456–457 method, 457–459, 458f troubleshooting, 459 protocols microinjecting holo-aequorin into dechorionated and intact zebrafish embryos, 197–205 dechorionating embryos, 200 discussion, 203 imaging chamber preparation, 198–200, 200f materials, 197–198 method, 198–202 micropipette preparation, 198, 199f recipes, 205 troubleshooting, 202–203 reconstitution of holo-aequorin with apoaequorin mRNA and coelenterazine in zebrafish embryos, 206–210 discussion, 209 materials, 206–207 method, 207–209 recipes, 210 troubleshooting, 209 using targeted variants of aequorin to measure Ca2þ levels in intracellular organelles, 185–191 calibration, 187, 188f discussion, 191 materials, 185–187, 186f measurement of Ca2þ in ER and its release, 190 measurement of capacitative Ca2þ influx, 188–189 measurement of cytosolic Ca2þ, 187–188 measurement of mitochondrial Ca2þ uptake, 189, 189f measurement of mitochondrial Ca2þ uptake in permeabilized cells, 190 recipes, 191 reconstitution of functional photoprotein, 187 transfection, 187 troubleshooting, 190–191 reduction of affinity to measure high [Ca2þ], 180 semisynthetic, 193 structure and function, 178–179, 178f, 193 targeting of, 179–180, 181t–182t Aequorin buffer (recipe), 204 Affinity of indicators for Ca2þ, 28, 533–534 of synthetic Ca2þ buffers, 537t Agarose, reactive red (RRA), 345 Agarose solution for building agar salt bridges (recipe), 353 Agatoxin IVA, 144 Airy rings, 6 Alcohol dehydrogenase, 495 Alkaline phosphatase, 527 Allen and Blinks algorithm, 187 Alzheimer’s disease, 323 American Type Culture Collection (ATCC), 445, 575 2-aminoethoxydiphenylborat (2-APB), 279, 287 Amiodarone, 451 AMP-5, affinity for Ca2þ, 537t Amphotericin-B, 280 Animals, humane treatment of, 573 Antimycin, 215 AP5, 144 2-APB (2-aminoethoxydiphenylborat), 279, 287 Apoaequorin, 193, 194, 206–209 Apoptosis, 451–452 AquaLite aequorin, 203 Arabidopsis high-resolution imaging of cytoplasmic Ca2þ dynamics in Arabidopsis roots (protocol), 474–478 discussion, 477 materials, 474–475 method, 475–477, 476f data analysis, 477 imaging, 475–477 plant growth, 475 sample preparation, 475, 476f troubleshooting, 477 imaging of mitochondrial and nuclear Ca2þ dynamics in Arabidopsis roots (protocol), 484–488 discussion, 487 materials, 484–485 method, 485–486 CLSM analysis, 486 imaging analyses, 486 sample preparation, 486 seed sterilization and growth of plantlets, 485–486 recipes, 487 troubleshooting, 487 live cell imaging of cytoplasmic and nuclear Ca2þ dynamics in Arabidopsis roots (protocol), 479–483 discussion, 482 materials, 479–480 method, 480–482, 481f Ca2þ imaging, 480 data analyses, 482 plant growth, 480 sample preparation, 480, 481f recipes, 483 troubleshooting, 482 live cell imaging of cytoplasmic Ca2þ dynamics in Arabidopsis guard cells (protocol), 469–473 discussion, 473 materials, 469–470 method, 470–472, 471f data analysis, 472 imaging, 471–472 plant growth, 470 sample preparation, 470–471, 471f recipe, 473 troubleshooting, 472–473 Artificial seawater (ASW) (recipe), 515, 523 Aspiration system, 38 Astrocytes, intercellular Ca2þ waves in, 95, 96 ATCC (American Type Culture Collection), 445, 575 ATP, 188 affinity for Ca2þ, 537t caged, 75, 111 intercellular Ca2þ waves (ICWs) and, 95 ATPases. See also Calcium (Ca2þ) pump calculation of activity, 238, 239f plasma membrane Ca2þ ATPase (PMCA), 231, 277, 291, 293, 438, 446 secretory pathway Ca2þATPase (SPCA), 231, 446 SERCA (see Sarco-endoplasmic Ca2þ ATPases) in yeast, 446 ATP-regenerating system, 513, 522 ATP stock solution for ER sensor calibrations (recipe), 63 Autoclave safety, 572 Auto-fluorescence, in two-photon microscopy, 16