One day after arrival, rats were measured for lean and fat body mass using nuclear magnetic resonance (Minispec LF90 Body Composition Analyzer; Bruker, East Milton, ON, Canada). Obese-prone rats were matched for body weights and randomly assigned to free feeding (Obese-FF) or food-restricted (Obese-FR) groups (six rats per group). Lean-prone animals were randomly assigned to free-feeding (Lean-FF) or food-restricted (Lean-FR) groups (six rats per group). In obese-prone rats, CR consisted of pair feeding the obese-prone rats the daily mean amount of food consumed by the Lean-FF group from the previous day. The daily food intake of Lean-FR rats was a percentage of the food consumption of Lean-FF animals; we calculated this percentage from the difference in daily food intake of Obese-FF and Obese-FR groups (Lean-FR intake=Lean-FF intake*(Obese-FF intake−Obese-FR intake)/Obese-FF intake*100). Thus, Obese-FR and Lean-FR groups were matched on relative average amount of CR. Animals were fed standard laboratory chow (LabDiet 5010 Rodent Diet, PMI Nutrition International Inc., Brentwood, MO, USA). An electronic scale (Sartoris, Model TE4101, Sartorius AG, Goettingen, Germany) was used to measure daily food and body weight to the nearest gram.
Minispec lf90 body composition analyzer
Manufactured by Bruker
Sourced in United States
The Minispec LF90 Body Composition Analyzer is a laboratory instrument designed to measure the body composition of samples. It utilizes low-field nuclear magnetic resonance (LF-NMR) technology to determine the total fat, water, and lean mass content of the sample.
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5 protocols using minispec lf90 body composition analyzer
1
Body Composition Analysis of Obese and Lean Rats
2
Gut Microbiome Modulation via FMT
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Antibiotic-treated mice with microbiota depletion were given daily oral gavage with a 200-µl antibiotics cocktail (ampicillin 1 g l−1, neomycin 1 g l−1, metronidazole 1 g l−1 and vancomycin 0.5 g l−1) for 7 d, followed by a 4-d antibiotic washout period before FMT. For FMT, the preparation of fresh stool samples and the subsequent operation in mice was conducted as previously described43 (link).
Body weight was measured every 3 or 4 d. Stool samples were collected before and after faecal transplantation and instantly stored at −80 °C until further analysis. Body composition was determined with a Minispec LF90 Body Composition Analyzer (Bruker). In vivo gut permeability, GTTs, ITTs and whole-body oxygen consumption were accessed via a comprehensive laboratory animal monitoring system (Columbus Instruments)68 (link). Fat mass in inguinal subcutaneous, epididymal, peri-renal and mesenteric white adipose tissue was determined after death by wet weight measurements. Blood and various tissues were collected for further biochemical evaluations. The study did not blind investigators to group allocations and no mice were excluded.
Body weight was measured every 3 or 4 d. Stool samples were collected before and after faecal transplantation and instantly stored at −80 °C until further analysis. Body composition was determined with a Minispec LF90 Body Composition Analyzer (Bruker). In vivo gut permeability, GTTs, ITTs and whole-body oxygen consumption were accessed via a comprehensive laboratory animal monitoring system (Columbus Instruments)68 (link). Fat mass in inguinal subcutaneous, epididymal, peri-renal and mesenteric white adipose tissue was determined after death by wet weight measurements. Blood and various tissues were collected for further biochemical evaluations. The study did not blind investigators to group allocations and no mice were excluded.
3
Induction of Obesity and P. gingivalis Infection in Mice
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All animal experiments were approved by the Committee for the Care and Use of Laboratory Animals at Fudan University. C57BL/6 male mice (8 weeks old, Beijing Vital River Laboratory Animal Technology Co., Ltd., Beijing, China) were group-housed in a specific pathogen-free (SPF) controlled environment with free access to food and water under a strict 12 h light/dark cycle. Forty mice were randomized into four equal groups with ten mice each: ND (normal diet), HFD (high-fat diet), sham, and Pg.
For the obese model, mice were fed a high-fat diet (HFD, 60% fat, 20% protein, and 20% carbohydrates, Research Diets, D12492) to induce obesity for 12 weeks, while a normal diet (ND, 10% fat, 20% protein, and 70% carbohydrates, Research Diets, D12450J) was used as a control. For P. gingivalis administration, the mice were gavaged with 109 CFU P. gingivalis twice a week for 6 weeks, and PBS with 2% carboxymethylcellulose was administered as a sham. Body weight was assessed in the last week, and fat mass was detected using a Minispec LF90 body composition analyzer (Bruker, Massachusetts, USA).
For the obese model, mice were fed a high-fat diet (HFD, 60% fat, 20% protein, and 20% carbohydrates, Research Diets, D12492) to induce obesity for 12 weeks, while a normal diet (ND, 10% fat, 20% protein, and 70% carbohydrates, Research Diets, D12450J) was used as a control. For P. gingivalis administration, the mice were gavaged with 109 CFU P. gingivalis twice a week for 6 weeks, and PBS with 2% carboxymethylcellulose was administered as a sham. Body weight was assessed in the last week, and fat mass was detected using a Minispec LF90 body composition analyzer (Bruker, Massachusetts, USA).
4
Dietary Effects on Metabolic Phenotypes
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The mice were randomly fed with low fat laboratory standard diet (SD) [composed of 20% protein, 5% fat, 4.7% fiber and 52.9% carbohydrate; 3.41 kcal/g; LabDiet, 5053 (LabDiet; St. Louis, MO, USA)] or high fat (HF) diet [composed of 24% protein, 24% fat, 5.8% fiber and 41% carbohydrate; 4.73 kcal/g; D12451 (Research Diet, New Brunswick, NJ, USA)] respectively. The consumption of diets and the gain of body weight were examined every week. At the age of 13-week, the mice were further divided into two subgroups and exposed to either air (2 groups) or intermittent hypoxia (IH; 2 groups) for four weeks (n = 6 for each group). The IH protocol had been described in our previous study [19 ]. During the air/IH exposure, the mice were continued their SD or HF diet respectively. Total fat mass and total lean mass were assessed by the Minispec LF90 Body Composition Analyzer (Bruker, Billerica, MA, USA) before sacrifice. Mice were sacrificed with overdose of sodium pentobarbital anesthesia (100 mg/kg, i.p.) after fasting overnight. Serum samples and liver tissues were collected, frozen in liquid nitrogen before keeping in −80 °C freezer for further use.
5
Whole Body Composition and Metabolism
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Whole body composition was measured in conscious lightly restrained mice using the minispec LF90 Body Composition Analyzer (Bruker Optics, Inc.). This minispec analyzer is based on time domain nuclear magnetic resonance spectroscopy and provides amounts of fat, lean tissue and free body fluid. Body composition was measured at four different time points during the study: before HED (‐7 weeks), before ASO treatment start (week 0), and following 3‐ and 6‐weeks of ASO treatment. In dose response studies, body composition was measured after 2‐weeks of ASO treatment. Metabolic rate was measured following 6–7 weeks of ASO treatment using an 8‐cage custom indirect calorimetry system (TSE Systems, Inc.). Each individual cage was equipped with infrared beams to detect ambulation, rearing movement, and fine motor movement along the x, y, and z axes. Inlet and outlet ports into each cage monitored O2 and CO2 levels, enabling calculation of oxygen consumption (VO2, ml/h), carbon dioxide production (VCO2, ml/h), and respiratory exchange ratio (RER). RER was calculated as the ratio VCO2/VO2, while energy expenditure was calculated as caloric expenditure through heat (kcal/h/kg bw). Mice were given 5‐days to acclimate to metabolic cages prior to 48‐h calorimetry measurement (last 24‐h used for analysis). A total of 12 animals were divided into two cohorts of six mice for calorimetry studies.
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