Experiment 2 FORMAL REPORT Draft

Cell and Molecular Biology Laboratory First Term Term 2016-2017

Exeriment 2 Cell !taining Tec"ni#ues Belen$ %lexis

Bitera$ &anicaBuena'entura$ (elsie Cantre$ (od)in Castillo$ *amila  Department of Biological Sciences, College of Science University of Santo Tomas, Espana Manila 1051

&ate !ubmitted+ ,ctober $ 2016

Introduction

Procedure

1. 10

µ

L o/ aramecium s. susension culture )as laced in t"e center o/ a glass slide

and a co'er sli )as lo)ered gently abo'e$ a'oiding traed bubbles. 2. % micros microscoe coe )as /ocuse /ocused d using t"e scanner scanner 3 3 and its lig"t lig"t source source )as ad4usted ad4usted to reduce glare. 5. T"e ob4ecti' ob4ecti'es es )ere )ere s"i/ted s"i/ted to to L, to see 'ery motile motile cells. cells. . %/ter)ards$ %/ter)ards$ a stri stri o/ /ilt /ilter er aer )as )as inserted inserted on one side side o/ t"e co'er co'er sli to to dra) out some excess li#uid. . T"e sides o/ o/ t"e co'er sli sli )ere coated coated )it" )it" etroleum etroleum 4elly using using a toot"ic toot"ic to re'ent re'ent t"e slide /rom drying out. 6. T"e ob4ecti'es ob4ecti'es )ere )ere t"en s)itc"e s)itc"ed d to 8, to identi/y identi/y t"e t"e cell structur structures es t"at can can be seen in t"e unstained cells. 7. %ll o/ t"e stes stes )ere reeat reeated ed to add Lugol9 Lugol9ss iodine$ iodine$ met"yl met"yl green green and nigrosin nigrosinee but at t"e ste )"ere )"ile t"e )ater )as dra)n out /rom one side )it" a /ilter aer sli$ a dro o/ t"e c"osen stain )as laced on t"e ot"er side.

Results and Discussion

Cell staining is a basic rocess in studying microscoic organisms. :n t"is rocess$ seci/ic arts o/ t"e cells and ot"er comonents o/ t"e cells are being s"o)n deending on t"e stain used and t"e tyes o/ cell staining tec"ni#ue t"at )as emloyed. !tains "a'e di//erent mec"anisms in maring t"e cell. :t in'ol'es seci/icity and ermeability.

Cell and Molecular Biology Laboratory First Term 2016-2017 aramecium s. is a unicellular ciliated roto;oan$ )"ic" is commonly used as reresentati'e o/ ciliates in s"o)ing t"e mo'ement and mec"anism o/ cilia. T"ey belong to  "ylum %l'eolata$ sub"ylum Cilio"ora and class nstained aramecium under L,

:mage2.0 >nstained aramecium under 8,

Cell and Molecular Biology Laboratory First Term 2016-2017 !e'eral dyes are being used in 'i'o imortant cell comonents and arts. ,ne o/ t"e stains t"at are being used in 'i'o is t"e Lugol9s iodine. Lugol9s iodine or Lugol9s solution is commonly used in laboratory in detecting t"e resence o/ olysacc"aride etru;;i$ et. al.$ 20103. :ts c"emical name is otassium triiodide.

Figure 1.0. C"emical structure o/ Lugol9s iodine or otassium iodide

T"e stain is also being used in oral cancer detection. :t is comosed o/ iodine and  otassium iodide dissol'ed in distilled )ater. :n t"e resence o/ amylose$ a solution colors )it" dee blue. ?"ile in t"e resence o/ amyloectin$ it gi'es a urle color$ )"ic" is t"e original color o/ iodine. :n t"e resence o/ glycogen$ )"ic" is resent in t"e animal cell as storage o/  glucose$ it gi'es a reddis" bro)n to bro)n blac color. T"e Lugol9s solution is being taen u by t"e glycogen /ound in t"e cell Bas/ord$ et.al.$20153. T"e iodine /orms comlexes )it" t"e linear  c"ain o/ t"e glycogen ia$ et.al.$ 2013.T"roug" t"is stain$ t"e cilia$ )"ic" are t"e locomotory organelle o/ t"e microorganism$ became 'isible and aear as reddis" bro)n color. T"is is due to t"e resence o/ glycogen t"at ser'es as energy storage$ ro'iding t"e re#uired energy /or t"e locomotory organelle. :n t"e exeriment$ since t"e microorganism is surrounded by t"eir cilia$ t"e )"ole organism )as stained )it" t"e Lugol9s iodine.

Cell and Molecular Biology Laboratory First Term 2016-2017

m:

:mage 5.0 aramecium cells stained )it" Lugol9s iodine under 8,

Met"yl green or met"ylene green is a basic or cationic dye t"at is being used in staining c"romatin &memura$ et. al.$ 20053. :t is a "eterocyclic aromatic comound t"at bears a  ositi'e c"arge in its amino grou Figure 2.03.

Figure 2.0. C"emical !tructure o/ met"yl green

Cell and Molecular Biology Laboratory First Term 2016-2017

Figure 5.0. C"emical !tructure o/ &