{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,25]],"date-time":"2025-09-25T15:40:54Z","timestamp":1758814854463},"reference-count":44,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2005,12,1]],"date-time":"2005-12-01T00:00:00Z","timestamp":1133395200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Inher Metab Disea"],"published-print":{"date-parts":[[2005,12]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Cytoplasmic granules in fibroblasts, visualized without stains, or labelled with Nile red, Filipin, or anti\u2010LAMP\u20101 (lysosome\u2010associated membrane protein 1), were imaged using the real\u2010time microscope (RTM). New advances in light microscope technology were applied to detect cytoplasmic granules (RTM\u2010visible granules) and characterize them by imaging contrast, size, shape, cellular distribution, composition, motion dynamics and quantity. Appearing as solid spheroids or ring structures, the majority of the RTM\u2010visible granules contained Nile\u2010red labelled neutral lipids. A smaller subpopulation, appearing dimmer, with less imaging contrast, contained Filipin\u2010labelled free cholesterol. Most lipid storage granules have a diameter ranging from 0.3 \u03bcm to 0.6 \u03bcm, with a small population measuring up to 1 \u03bcm. They typically clustered in the perinuclear region and displayed relatively small oscillatory motion. Immunofluorescence based on LAMP\u20101 labelling highlighted granular structures that were distinct and separate from RTM\u2010visible granules and other structures in the light modality of the microscope. RTM\u2010visible granules were associated with disease phenotypes that have increased cellular neutral lipid stores corresponding to the Nile red\u2010labelled droplets (e.g. triacylglycerides, cholesterol esters). As predicted, the fibroblast strains with a defect resulting in Wolman disease, when compared to control samples, consistently had RTM\u2010visible granules, higher in imaging contrast and with larger diameters, that were labelled with Nile red, and also an increased frequency of Filipin\u2013cholesterol complexes. By comparison, in fibroblasts where the lipid storage is less evident (Gaucher and Farber diseases) or from GM<jats:sub>1<\/jats:sub> gangliosidosis, where the primary storage substances are oligosaccharides, fewer and smaller RTM\u2010visible granules were observed. In some cases, changes in contrast and morphology in the unstained cytoplasmic compartments were more evident than in the labelled structures. In summary, applying the RTM imaging system to fibroblasts enables differences between the various disease types to be seen and, in specific examples, a unique phenotype can be readily discerned.<\/jats:p>","DOI":"10.1007\/s10545-005-0117-7","type":"journal-article","created":{"date-parts":[[2006,1,25]],"date-time":"2006-01-25T09:35:29Z","timestamp":1138181729000},"page":"991-1004","source":"Crossref","is-referenced-by-count":13,"title":["A comparative study of cytoplasmic granules imaged by the real\u2010time microscope, Nile Red and Filipin in fibroblasts from patients with lipid storage diseases"],"prefix":"10.1002","volume":"28","author":[{"given":"N.\u2010A.","family":"Pham","sequence":"first","affiliation":[{"name":"Richardson Technologies Inc. Toronto Ontario"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. R.","family":"Gal","sequence":"additional","affiliation":[{"name":"Richardson Technologies Inc. Toronto Ontario"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. D.","family":"Bagshaw","sequence":"additional","affiliation":[{"name":"The Hospital for Sick Children Research Institute Elm Wing, Room 9144, 555 University Avenue Toronto M5G 1X8 Canada"},{"name":"Department of Laboratory Medicine and Pathobiology University of Toronto Toronto Ontario Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A. J.","family":"Mohr","sequence":"additional","affiliation":[{"name":"Richardson Technologies Inc. Toronto Ontario"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B.","family":"Chue","sequence":"additional","affiliation":[{"name":"Richardson Technologies Inc. Toronto Ontario"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T.","family":"Richardson","sequence":"additional","affiliation":[{"name":"Richardson Technologies Inc. Toronto Ontario"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J. W.","family":"Callahan","sequence":"additional","affiliation":[{"name":"The Hospital for Sick Children Research Institute Elm Wing, Room 9144, 555 University Avenue Toronto M5G 1X8 Canada"},{"name":"Department of Biochemistry University of Toronto Toronto Ontario Canada"},{"name":"Department of Pediatrics University of Toronto Toronto Ontario Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,12]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bjc.6600188"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajplung.1999.277.4.L848"},{"key":"e_1_2_1_4_1","first-page":"1861","article-title":"The relationship between nuclear magnetic resonance\u2010visible lipids, lipid droplets, and cell proliferation in cultured C6 cells","volume":"59","author":"Barba I","year":"1999","journal-title":"Cancer Res"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399-0004.1989.tb03364.x"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.240480113"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.85.21.8022"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1177\/35.5.3559182"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/jnr.490270411"},{"key":"e_1_2_1_10_1","first-page":"295","article-title":"Use of Nile red stain in the detection of cholesteryl ester accumulation in acid lipase\u2010deficient fibroblasts","volume":"112","author":"Brown WJ","year":"1988","journal-title":"Arch Pathol Lab Med"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj2850833"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/28.4.569"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1177\/33.8.4020099"},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.45.2.334"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00418-002-0462-7"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2275(20)32376-2"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2275(20)34307-8"},{"key":"e_1_2_1_18_1","first-page":"195","article-title":"Quantitative analysis of the movements of cytoplasmic granules in polarized fibroblasts","volume":"11","author":"Grigoriev IS","year":"1997","journal-title":"Membr Cell Biol"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.100.5.1488"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1177\/002215549704500513"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/10084"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/0021-9150(90)90109-V"},{"key":"e_1_2_1_23_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.83.3.588"},{"key":"e_1_2_1_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2275(20)39846-1"},{"key":"e_1_2_1_25_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.157.3790.804"},{"key":"e_1_2_1_26_1","first-page":"323","article-title":"Electron microscopic studies in lipid storage disease","volume":"17","author":"Ludatscher RM","year":"1981","journal-title":"Isr J Med Sci"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.48.32304"},{"key":"e_1_2_1_28_1","doi-asserted-by":"publisher","DOI":"10.1017\/S143192760404005X"},{"key":"e_1_2_1_29_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00273194"},{"key":"e_1_2_1_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/2281093b0"},{"key":"e_1_2_1_31_1","doi-asserted-by":"publisher","DOI":"10.5694\/j.1326-5377.1980.tb100764.x"},{"key":"e_1_2_1_32_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.113.17.2977"},{"key":"e_1_2_1_33_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1768-322X.1988.tb00704.x"},{"key":"e_1_2_1_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0344-0338(11)80422-X"},{"key":"e_1_2_1_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)69086-6"},{"key":"e_1_2_1_36_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M211289200"},{"key":"e_1_2_1_37_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/10.2.127"},{"key":"e_1_2_1_38_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF01800716"},{"key":"e_1_2_1_39_1","doi-asserted-by":"publisher","DOI":"10.1089\/10430340150218413"},{"key":"e_1_2_1_40_1","doi-asserted-by":"publisher","DOI":"10.1002\/1531-8257(200011)15:6<1199::AID-MDS1020>3.0.CO;2-1"},{"key":"e_1_2_1_41_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1022575511354"},{"key":"e_1_2_1_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/0925-4439(91)90069-L"},{"key":"e_1_2_1_43_1","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2760(79)90035-3"},{"key":"e_1_2_1_44_1","doi-asserted-by":"publisher","DOI":"10.1006\/excr.1999.4774"},{"key":"e_1_2_1_45_1","doi-asserted-by":"publisher","DOI":"10.1042\/0264-6021:3480621"}],"container-title":["Journal of Inherited Metabolic Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1007\/s10545-005-0117-7","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1007\/s10545-005-0117-7","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,15]],"date-time":"2023-09-15T02:15:58Z","timestamp":1694744158000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1007\/s10545-005-0117-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,12]]},"references-count":44,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2005,12]]}},"alternative-id":["10.1007\/s10545-005-0117-7"],"URL":"https:\/\/doi.org\/10.1007\/s10545-005-0117-7","archive":["Portico"],"relation":{},"ISSN":["0141-8955","1573-2665"],"issn-type":[{"value":"0141-8955","type":"print"},{"value":"1573-2665","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,12]]}}}