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The sections were then incubated with Alexa fluor 555-labeled secondary antibody (1:1000 in 1x PBS pH 7.4) (Invitrogen, Thermo Fisher Scientific) for 1 hour at room temperature in a dark, humidified chamber, followed by washing in PBS

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If follow-up has lapsed which is not uncommon several years after surgery patients are encouraged to re-engage with their GP to arrange a nutritional review and, where appropriate, re-referral to the original bariatric unit or a local specialist service