Assessment of different protocols for the isolation and purification of gut associated lymphoid cells from the gilthead seabream (Sparus aurata L.)

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Author: Salinas, I
Author: Esteban, M
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Methods Article

Assessment of different protocols for the isolation and purification of gut associated lymphoid cells from the gilthead seabream (Sparus aurata L.)

Irene Salinas¹, José Meseguer¹ and Maria Ángeles Esteban¹^*

¹ Fish Innate Immune System Group, Department of Cell Biology, Faculty of Biology, University of Murcia. 30100 Murcia. Spain. Phone: +34 968367665 Fax: +34 968363963 Email: aesteban@um.es.

* To whom correspondence should be addressed: Maria Ángeles Esteban, Fish Innate Immune System Group, Department of Cell Biology, Faculty of Biology, University of Murcia. 30100 Murcia. Spain. Phone: +34 968367665. Fax: +34 968363963. Email: aesteban@um.es

Biol. Proced. Online 2007;9:43-55. doi:10.1251/bpo132
Submitted: February 15, 2007; Accepted: March 30, 2007; Published: November 25, 2007.

Indexing terms: Sea Bream; Flow Cytometry; Respiratory Burst.

Figure 4 Enlarged

Fig. 4:

Light micrographs of semithin sections of seabream intestine at different stages of cell extraction. L (lumen), E (enterocytes), LP (lamina propria). (a) Gut semithin section at the beginning of the isolation procedure (control sample, time 0). Bar: 10 μm. (b) Gut semithin section after 10min in DTT. Bar: 10 μm (c) Gut semithin section after 10 min in DTT and 60 min in collagenase (0.15 mg/ml). Note the open interepithelial spaces (arrows) that allow leucocytes to be freed into the media. Bar: 10 μm.

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