Digesta collected from the distal jejunum were divided into two tubes (15 mL) and centrifuged at 1,000 × g at 4 °C for 10 min. The supernatant of each tube was collected into 2 mL tubes and centrifuged at 10,000 × g at 4 °C for 10 min. The supernatant was then transferred to a new 2-mL tube and kept on ice for the measurement of viscosity. A viscometer (Brookfield Digital Viscometer, Model DV-II Version 2.0, Brookfield Engineering Laboratories Inc., Stoughton, MA) was then used to measure the digesta viscosity at 25 °C. The viscosity was calculated as the average between 45.0/s and 22.5/s shear rates and recorded as millipascal-seconds (mPa.s) (Duarte et al., 2019 (link); Chen et al., 2020 (link)).
Brookfield digital viscometer model dv 2 version 2
Manufactured by Ametek
The Brookfield Digital Viscometer, Model DV-II Version 2.0 is a laboratory instrument designed to measure the viscosity of fluids. It uses a rotating spindle immersed in the sample to determine the sample's resistance to flow, providing a quantitative measure of its viscosity.
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5 protocols using brookfield digital viscometer model dv 2 version 2
1
Viscosity Analysis of Digesta from Distal Jejunum
2
Measurement of Intestinal Digesta Viscosity
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Following the procedure by Passos et al. (2015) (link) and Duarte et al. (2019) (link), digesta collected from the mid-jejunum was divided into two tubes (15 mL) and centrifuged at 1,000 × g for 10 min at 4 °C to obtain the liquid phase. The supernatant of each tube was collected into 2 mL tubes and centrifuged at 10,000 × g at 4 °C for 10 min. The supernatant obtained was transferred to another 2 mL tube and kept on ice for further assessment of viscosity. The amount of 0.5 mL of digesta supernatant was placed in the viscometer (Brookfield Digital Viscometer, Model DV-II Version 2.0, Brookfield Engineering Laboratories Inc., Stoughton, MA) set at 25 °C. The viscosity measurement was the average between 45.0 and 22.5 per s shear rates, and the viscosity values were recorded as apparent viscosity in millipascal-seconds (mPa.s).
3
Measurement of Jejunal Digesta Viscosity
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Samples of jejunum digesta from 50 mL tubes were divided into 2 falcon tubes (15 mL) and centrifuged at 1,000 × g at 4 °C for 10 min to obtain the liquid phase. Then, the liquid phase was removed to an Eppendorf tube (2 mL) to centrifuge at 10,000 × g at 4 °C for 10 min. The supernatant obtained was transferred to another Eppendorf tube (1.5 mL) for further measurement. 0.5 mL of digesta supernatant was placed in the viscometer (Brookfield Digital Viscometer, Model DV-II Version 2.0, Brookfield Engineering Laboratories Inc., Stoughton, MA), set at 25 °C. The viscosity measurement was the average between 45.0/s and 22.5/s shear rates, and the viscosity values were recorded as apparent viscosity in millipascal-seconds (mPa s).
The pH was measured immediately after collecting using a pH meter (AB15 Basic, Thermo Fisher Scientific Inc., Waltham, MA), placed directly in the digesta.
The pH was measured immediately after collecting using a pH meter (AB15 Basic, Thermo Fisher Scientific Inc., Waltham, MA), placed directly in the digesta.
4
Measurement of Jejunal Digesta Viscosity
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Viscosity was immediately measured in the same day after obtaining jejunal digesta based on the methods described by Passos et al. (2015b) (link). The jejunal digesta were homogenized and then transferred to 15 mL tubes (each sample repeated four times). The tubes were centrifuged at 1,500 × g for 5 min at 4 °C and then the supernatant was again centrifuged at 10,000 × g for 10 min at 4 °C. The second supernatant was placed in ice to measure viscosity immediately. A viscometer (Brookfield Digital Viscometer, Model DV-II Version 2.0, Brookfield Engineering Laboratories, Inc.) was set at 25 °C, and 1.0 mL of the supernatant was placed in the viscometer.
5
Jejunum Digesta Viscosity Measurement
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Following the procedure by Passos et al. (2015) (link) and Duarte et al. (2019) (link), samples of jejunum digesta from 50 mL tubes were divided into 2 falcon tubes (15 mL) and centrifuged at 1,000 × g at 4 °C for 10 min to obtain the liquid phase. The liquid phase was then removed and transferred to an Eppendorf tube (2 mL) to centrifuge at 10,000 × g at 4 °C for 10 min. The supernatant obtained was transferred to another Eppendorf tube (1.5 mL) for further measurement. About 0.5 mL of digesta supernatant was placed in the viscometer (Brookfield Digital Viscometer, Model DV-II Version 2.0, Brookfield Engineering Laboratories Inc., Stoughton, MA), set at 25 °C. The viscosity measurement was the average between 45.0 s−1 and 22.5 s−1 shear rates, and the viscosity values were recorded as apparent viscosity in centipoise (cP ).
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