[Leish-l] Fwd: Growth curve in promastigote culture

[Gabriel Ferreira]

Dear Nayrone,


As you likely know, there are various methods to perform parasite growth
curves. I've used both plaque assays and flask cultures. Plaque assays
offer the advantage of assessing multiple strains simultaneously. Optic
densitometry (OD) can be employed if it's available in your lab. If you
want to do it in flask cultures, I typically start with a 5 mL SDM medium
culture containing 10e6/mL (5x10e6 total) of healthy promastigotes in the
log-phase. Daily parasite counts are then performed, usually up to day 6 or
adjusted according to experiment goals. During initial days, 1:50 dilution
works well, shifting to 1:100 afterward to avoid low parasite numbers at
the start and excessive numbers at the end of the cycle. For those less
experienced in visualizing *Leishmania *promastigotes, diluting the culture
in PBS-glycerol can reduce parasite mobility.



Hope this can be of any help!





Best wishes


Gabriel

Em ter., 15 de ago. de 2023 às 09:13, Carlos Costa <chncosta at gmail.com>
escreveu:

> Gabriela,
>
> Ninguém melhor do que você para responder a esta jovem pesquisadora.
>
> Abraços,
>
> Carlos.
>
> ---------- Forwarded message ---------
> From: Nayore Tk <nayoretk at gmail.com>
> Date: Mon, Aug 14, 2023 at 12:40 PM
> Subject: [Leish-l] Growth curve in promastigote culture
> To: <leish-l at lineu.icb.usp.br>
>
>
> Good afternoon,
>
> My name is Nayore, I am a doctoral student and researcher at UFSCar and I
> work with Leishmania infantum promastigotes culture. I have doubts about
> how to perform the growth curve, should I do it in 12 well plate or in a
> culture flask?
>
> I performed this type of count for other strains, however I was performing
> the plate count during 10 days, starting from a concentration of 2.0x10e5
> promastigotes/mL in first day. Recently I was asked why I was making it on
> a well and not in a flask.
>
> I would like to solve this question about which is the best way to make the
> curve in a well containing 2.0x10e5 promastigotes/2 mL or in a culture
> flask?
>
> thank you for your attention
>
> Best Regards
> Nayore Tamie Takamiya
>
> --
> *Nayore Tamie Takamiya*
> Bióloga/ Doutoranda no Programa de Pós-graduação em Genética Evolutiva e
> Biologia Molecular
> Universidade Federal de São Carlos (UFSCar)
> Centro de Ciências Biológicas e da Saúde (CCBS)
> Departamento de Genética e Evolução (DGE)
> Rod. Washington Luís, Km 235 Caixa Postal 676
> CEP. 13565-905 - São Carlos - SP - Brasil
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>
> --
> Carlos H. N. Costa, MD, DSc.
> Head
> Instituto de Doenças do Sertão
> *Instituto de Doenças Tropicais Natan Portella*
>
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-- 
--
Gabriel Reis Ferreira, BMS, MSc.
PhD student
Université LavaL
Département de microbiologie-infectiologie et d'immunologie.

Division of Infectious Disease and Immunity

CHU de Québec Research Center-Laval University

2705 Laurier Blvd. Quebec, QC, Canada G1V 4G2

Tel: 1-(418) 525-4444
Ulaval e-mail: gabriel.reis-ferreira.1 at ulaval.ca
<gabriel.reis-ferreira.1 at ulaval.ca>
CHUL e-mail: gabriel.ferreira at crchudequebec.ulaval.ca
<gabriel.ferreira at crchudequebec.ulaval.ca>