Use of a carbocyanine dye as a marker of functional vasculature in murine tumours

Trotter, M.J.; Dj, Chaplin; Olive, Peggy L.

doi:10.1038/bjc.1989.148

Cited by 81 publications

(50 citation statements)

References 12 publications

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“…The rapid clearing of lectin from circulating blood is comparable to rapid clearance of other dyes (Trotter et al 1989) and also with other reports that intravascular injection of Evans blue results in generalized body staining within 15 s (Debbage et al 1998). Clearance from blood is probably due to a combination of endothelial binding, uptake by the reticulo-endothelial system, some tissue extravasation, and renal clearance.…”

Section: Temporal Aspects Of Tomato Lectin Bindingsupporting

confidence: 83%

Use of labeled tomato lectin for imaging vasculature structures

Robertson

Levine

Haynes

et al. 2014

Histochem Cell Biol

154

118

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Section: Temporal Aspects Of Tomato Lectin Bindingsupporting

confidence: 83%

Use of labeled tomato lectin for imaging vasculature structures

Robertson

Levine

Haynes

et al. 2014

Histochem Cell Biol

154

118

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“…This technique is very sensitive to focal, complete loss of blood flow because it compares the pre-and post-treatment perfusion patterns in the same tissue section (Zwi et al, 1989). H33342 is a useful first vascular marker because of its low toxicity, diffusion characteristics and stability in vivo (Trotter et al, 1989). However, in our hands, DiOC7(3) showed acute toxicity at doses required for adequate tissue fluorescence, and showed different diffusion properties to those of H33342 (see below).…”

Section: Discussioncontrasting

confidence: 42%

“…The fluorescent marker study used was based on that of Trotter et al (1989), in which each of two fluorescent dyes, H33342 and the carbocyanin dye DiOC7(3), are injected intravenously at different times. These dyes stain the tumour tissue close to those vessels which are functional at the time of injection, allowing changes in perfusion status of individual vessels to be observed.…”

Section: Discussionmentioning

confidence: 99%

“…To determine the role of the vasculature in the anti-tumour action of FAA, we have examined the process of attachment and vascularisation of EMT6 spheroids in the peritoneum, and found tumours which were directly comparable in site and size range to avascular spheroids (AVS). Since these vascularised spheroids (VS) Trotter et al (1989), was used to assess the changes in blood flow in the VS at 4 h.…”

mentioning

confidence: 99%

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The use of vascularised spheroids to investigate the action of flavone acetic acid on tumour blood vessels

Zwi

Bc²,

Gavin³

et al. 1990

Br J Cancer

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Summary EMT6 multicellular spheroids were introduced into the peritoneal cavities of mice and allowed to become vascularised, resulting in solid spherical tumours. The necrotic cores of the initially avascular spheroids were replaced by vascularised tumour tissue but the outer zones of the spheroids failed to become vascularised. The presence of both vascular and avascular components in each spheroid allowed the role of the vasculature in the antitumour action of flavone acetic acid (FAA) to be determined. Eighteen hours after treatment with FAA 0.8 mmol kg-', the vascularised core became necrotic and haemorrhagic, while the outer avascular zone remained viable. Tumour Materials and methodsHistological studies EMT6 multicellular tumour spheroids were grown in spinner flask culture in a-MEM + 10% fetal calf serum, and between 15 and 25 spheroids, varying in size from 0.5 to 1.2 mm in diameter, were injected into the peritoneal cavities of anaesthetised Balb/C mice through a 161 gauge needle, as described previously (Zwi et al., 1989). Seven days later the mice in the treatment group (n = 7) were injected with FAA 0.8 mmol kg-' (kindly supplied by Dr K.D. Paull, National Cancer Institute) in 5% w/v sodium bicarbonate by the intravenous (i.v.) or intraperitoneal (i.p.) route, and were killed after a further 18 h. Untreated spheroid-bearing mice (n = 2) were killed after the same interval. The peritoneal cavities of the mice were opened, the free spheroids were collected, and those spheroids which had become attached to host structures were excised with a cuff of adjacent tissue. The spheroids were fixed in either 4% neutral formaldehyde or 2.5% phosphate buffered glutaraldehyde (pH 7.4). The formalin-fixed tissue was embedded in paraffin, and 5 gm sections were stained with haematoxylin and eosin (H&E). The glutaraldehyde-fixed spheroids were embedded in epoxy resin and 2 gim sections stained with toluidine blue. Multiple sections were examined from each spheroid.

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“…Hoechst 33342 and DiOC7(3) have short distribution half-lives in blood and provide selective staining of tumour cells adjacent to functional vessels (Trotter et al, 1989b(Trotter et al, , 1990. The excitation and emission spectra differ between the dyes, allowing separate detection by fluorescence microscopy (Trotter et al, 1989a).…”

Section: Double-fluorescent Staining Techniquementioning

confidence: 99%