Cosmeyenn.

PART 1 / GRAMMAR

German tenses & moods 

Present tense 1 ; 2 ; 3 ( exercise: here ) 

Simple Past 1 ; 2 ; 3 ( here )

Present Perfect Tense 1 ; 2 ; 3 ( here )

Past Perfect Tense 1 ; 2 ; 3 ( here )

Future Tense 1 ; 2 ; 3 ( here )

Future Perfect Tense 1 ; 2 ; 3 ( here )

Imperative Mood 1 ; 2 ; 3 ( here )

Passive Voice 1 ; 2 ; 3 ( here )

Reflexive 1 ; 2 ; 3 ( here )

Subjunctive II 1 ; 2 ( here )

Subjunctive I 1 ; 2  ( here ) 

!! Conjugator for German Verbs !!

Nouns and Articles

Gender 

Articles

Plurals ( here )

Nominative

Accusative

Dative

Genitive

Pronouns

Personal Pronouns 1 ; 2 ; 3  ( here )

Possessive Pronouns 1 ; 2 ; 3  ( here )

Reflexive Pronouns 1 ; 2 ; 3 (  here )

Relative Pronouns 1 ; 2 ; 3   (  here )

Interrogative Pronouns 1 ; 2 ; 3  (  here )

Demonstrative Pronouns 1 ; 2 ; 3 ( here )

Indefinite Pronouns 1 ; 2  ( here )

Nominative 

Accusative

Dative

Genitive 

Adjectives

Types

Formation

Declension 

Comparative Form

exercises (with Adjectives):  Opposites ; Endings ;  Superlative ;  Comparative ;  Comparative and superlative ; Declension

youtube:  Adjectives & Opposites ;  Adjective Endings ;  Adjective Endings - Nominative - Definite and Indefinite Articles ;  Adjective Endings - Accusative - Definite and Indefinite Articles ;  Adjectives ; Basic German Adjectives; Declining German Adjectives 

Adverbs

Types

Location and Comparative Forms of Adverbs

exercises (with Adverbs):  Adverbs of place ;  Adverbs of time ;  Adverbs of manner

youtube:  Adverbs of Time; Adverbs of Time ;  Adverbs of Time ; 

Adverbs of Time  ; Adverbs of Manner ;  Adverbs of Place ;

Prepositions

Types

Cases

exercises (with Prepositions):  Dative prepositions ; Accusative prepositions ;

Genitive prepositions ;  Dative and accusative prepositions ; Prepositions of place ;  Prepositions of time  ;  Prepositions: aus or von  ;  Prepositions: nach or zu

youtube:  IN ; AN ; POSITIONSVERBEN ; AUF ; ÜBER ; UNTER ; ZWISCHEN ;  NEBEN - DANEBEN - NEBENEINANDER ;  HINTER - DAHINTER

Declension (in general)

Sentence Structure

Main Clauses 

Negation

Questions 

Dependent or Subordinate Clauses 

youtube:  Position of Main Verbs in Main Clauses ;  Questions ; Time Manner Place ;  Direct & Indirect Objects ;

Conjunctions

Introduction

Indirect Questions

Infinitive Clauses

Participle Clauses

Relative Clauses

Conditional Clauses

Indirect Speech

exercises(with Conjunctions): here ;  Subordinate conjunctions ; Conjunctions: dass or weil ; Conjunctions: dass or ob ;  Conjunctions: als, wenn, wann or ob ;  Conjunctions: als, wenn or wann

That’s all! GOOD LUCK!

Primary Human Epidermal Keratinocytes, adult (C-005-5C)

Thaw procedure

Reagents (do not warm in water bath):

50mL Epilife medium (M-EPI-500CA)

500uLHuman Keratinocytes Growth Supplement (S-005-1)

Steps

Prepare a 37C water bath for the thawing of cryopreserved primary cells.

Sterilize tissue culture hood with 75% alcohol.

Mix the two reagents (do not warm in water bath) in laminar flow culture hood and pour 7mL into a 10cm plate (Corning). Label with date, cell type, passage number, and your name.

Thaw the primary cells in 37C water bath with gentle agitation (around 1 to 2 minutes). Do not thaw thoroughly, a movable sliver of ice should still be seen in the cryogenic tube. Wipe cryogenic tube with 75% alcohol.

Transfer thawed primary cells into plate in laminar flow culture hood.

Use 500uL of fresh medium to wash cryogenic tube by pipetting up and down. Transfer medium into plate.

Store cell in a humidified 37C incubator with 5% CO^2, 95% air.

Do not disturb cell for at least 24 hours before changing cell media the next day by aspirating media then pouring in 7.5mL of fresh media.

They should have a cobblestone appearance, as shown in the picture below.

Maintenance procedure

Reagents (do not warm in water bath)

Maintenance medium (Epilife + HKGS)

Reminder

Change media every other day until 85% confluence.

Subculture procedure

Reagents

Trypsin/EDTA solution 0.025%

Maintenance media (Epilife medium + HKGS)

Trypsin Neutralizer Solution,TNS (R-002-100)

Steps

Thaw TNS in 37C water bath, and take out when thawed to cool off.

Sterilize tissue culture hood with 75% alcohol.

Move culture plate to laminar flow culture hood, aspirate medium.

Wash cells with 1mL trypsin/EDTA solution, remove solution after ensuring that entire surface was covered.

Add 1mL trypsin/EDTA and incubate at room temperature for approximately 8 to 10 minutes (check every 4 minutes) until the cells are completely round.

Check under microscope and lightly rap/tap under and on the sides of the the plate to dislodge cells.

When cells are detached, tilt the plate a little and add 3mL TNS into plate. “Wash” cells by gently pipetting solution over the plate several times.

Transfer solution into a sterile 15mL conical tube.

Add 3mL of fresh TNS into plate while tilting the plate. Repeat “wash” action, and transfer solution to the 15mL conical tube. A total of 7mL of solution should be in the 15mL conical tube.

Centrifuge cells at 180g (or 1268rpm for a 100mm radius motor) for 7 minutes.

Observe pellet, then remove supernatent gently from tube.

Resuspend cells gently in maintenance medium by pipetting up and down for a homogenous mixture.

Determine concentration of cells in suspension.

Seed new culture plate with at least 2.5x10^3cells/cm^2.

Incubate in humidified incubator at 37C, with 5% CO^2 and 95% air.

Cryopreserving procedure

Reagents

Trypsin/EDTA 0.025%

CryoStor CS10 Cell Crypreservation Media (C2874)

Maintenance medium (Epilife medium + HKGS)

Steps (if you would like to cryoperserve a tube and continue on with culturing)

Ensure 95% confluency of cells.

Follow “Subculture procedure” steps 1 to 9.

Transfer 4mL of solution to conical tube 2 and centrifuge both tubes at 180g (1268rpm for a 100mm radius motor).

Aspirate solution from both tubes.

Add 1mL to 2 mL cold CryoStor solution to conical tube 1 (previously with 3mL trypsin/EDTA/TNS solution) and resuspend pellet by gentle pipetting. Transfer solution to cryogenic tube and store in -80C overnight before transferring cryogenic tubes to liquid nitrogen.

Resuspend conical tube 2 (previously with 4mL trypsin/EDTA/TNS solution) with maintenance medium and add back to plate with fresh maintenance medium. Incubate plate in humidified incubator at 37C with 5% CO^2 and 95% air.

How was your mental health at first?

Some people (like my dad) assume that being a Ph.D. student is a breeze. The following are comments I hear most frequently:

“It’s only 3 years, right? And they even pay you. How nice of them!”

“It’s just another academic goal, it shouldn’t be harder than a bachelor’s degree.”

“You just need to hand in a few articles, how long does it take? 2 weeks?”

“You’re guaranteed a high-paying job when you graduate, what’s there to fuss about?”

And I’d always have to remind him:

“3 years, if I’m lucky. I pray that my data are on point during every experiment. The payment? It’s the scholarship. It wasn’t easy to acquire. But yes, I am very very grateful for it.”

“Yes, it’s my goal. Both are strenuous, for different levels of myself. I was a toddler back then, now I’m just a child in the academic world.”

“2 weeks? Probably for the reviewer to wonder whether or not my article is worth reviewing. 2 weeks is sometimes not even enough for one graph.”

“A gua- where did you get that idea?”

And that’s just part of it. At first, I would feel offended, but over time it clicked in me that he just didn’t understand my work.

Then I learned that it was okay to not be understood. I just have to educate him over time. Maybe one day he’ll get it.

      2.  How is your mental health now?

I’m going to be honest, it fluctuates every day. I could have a great day where I’d love to chat with everybody in the lab, then the next day I’d just rather be alone.

My professor, lab pals, friends, partner and family are amazing people, I’ve got nothing against them. But I always felt like it wasn’t enough, because I would always hold myself back when we’re conversing. There were just some problems I haven’t dealt with, because of tight schedules throughout the year (or life). 

I’m the kind of student who would:

feel ashamed of taking days off, even though I see my other lab members doing it consistently (and I was, of course, jealous for no reason). 

have a hard time rejecting work from my professor. At a point in life I was only sleeping 2 to 3 hours a day for a few months straight (I regret it now).

have a hard time correcting my juniors, even though it was for their own good. I didn’t want to make them unconfident in their work, their were just some tweaks that could be made.

It was horrible for the others and myself, and I wanted to change. 

So I went to counseling. And it was the best choice I could’ve ever made. It took half a year, but I improved myself to keep myself alive and human.

Now I’m able to:

ask for 1 to 2 days off from the lab, and just reboot my thoughts.

have better communication with my professor

convince my mind that what I’m about to say to my juniors are beneficially for their experiment.

Some background info, my family doesn’t help me financially because I’m already an adult. So my scholarship, and the extra income by working extra projects are saved for my school fee and life expenses.

Good day/afternoon/evening,

I’m here for a small introduction on why I started this blog and who I am.

Why start a blog?

Well, I’d first like to reconnect with my passion for writing!

Second, I know how it feels to go through articles and websites just to find that specific protocol for your experiment. Sometimes it doesn’t just take hours, it takes days, and you’ll be left feeling more lost than ever.

This blog should be for those who are looking for a THIS-COULD-MAYBE-BE-IT moment. I’m not saying these protocols are the one and only steps you should be taking, but they worked for me :)

Lastly, it would be nice to have people either commenting or reposting the protocols with the special twist that works for them!

Science should be sharable, cause most of us just want to learn and understand.

       2.  Who are you?

I’m currently a first year PhD student studying biomedical science. This field is my passion, yet the stress is literally sucking the life out of me.

Raccoon is the name I gave myself because…food is life, and the bed is my one and only true love… I have dark dark circles under my eyes, ok? If I don’t conceal them, my professor would think that I’m either sick or ready to transfer to the afterlife (one of them is true tho).

Things I’ll be posting:

Biomedical-related protocols

How my weekly schedule would look like on a chill/loaded week

Articles that I find interesting

Laboratory life

Mental health considerations

etc…

IMPORTANT NOTICE: I’m not always right, but I’ll always be open for discussions!

Drinking water

No longer making self deprecating jokes

Making sure to take time out of the day to relax and take a breather

Lighting candles

Counting ur breaths in and out if ur having a panic attack

Follow directions from your local/regional/national infectious disease prevention authorities.

DON’T PANIC: Even in the city of Wuhan, where the disease was first identified and there are the most verified cases (even if all 900+ cases in China were localized to Wuhan (they aren’t)) there are still less than 1 verified case per 10,000 people (as of Jan 24, 2020). Your relative risk of exposure is low, even if you are in that city.

Please correct people referring to this virus as “Wuhan Coronavirus” or “Wuhan Fish-market Coronavirus.” The proper name is 2019 NCoV (en-cahv or en-cove) or 2019 Novel Coronavirus. We don’t name viruses by the city or town that they were discovered in because of the economic harm that is done by permanently associating a disease with a specific location. We use broader geographic names or a naming system focused on year and type.

Wash your hands or use hand sanitizer regularly, but within reason (too much can dry your skin out.). Basic hygiene will decrease your risk of contracting or transmitting any number of diseases. And yes, guys, even if you only “went number 1,” wash your hands. Penis sweat mixes with taint sweat, mixes with butcrack sweat. Wash your hands.

If you live in a city with an active outbreak of a disease spread by droplets or airborne particles, feel free to wear a face mask/respirator rated at N-95. (Droplets are the visible particles released when you sneeze. Airborne means that when/if those droplets dry out, the virus or bacteria survive and remain infectious. Many diseases spread by droplet routes cannot survive drying.) I’m more likely to wear a mask if air quality is bad, but if it makes you feel safer, it’s ok.

THIS MAY BE THE SINGLE MOST IMPORTANT WAY TO PREVENT ILLNESS AND DEATH FROM DISEASES SPREAD BY AIRBORNE/DROPLET ROUTES.

GET YOUR DAMN FLU SHOT!

In the United States alone, every year, approximately 9.3-45 million people will have the flu. 140-810 thousand people will be hospitalized due to the flu, and 12-81 thousand people will die from the flu. (The wide range is due to some strains being more severe than others.) Beyond that, the risk of death from pneumonia and heart attacks/strokes goes up for the year after someone has a case of the flu, especially in elderly patients.

If you are around children, elderly people, people with weakened immune systems (cancer patients, organ transplant patients, etc.), or people that are not able to receive the flu shot because they are allergic to ingredients in the shot, or are unable to get the shot for any other reason, and you would rather not be responsible for increasing their risk of getting sick or dying, get your flu shot.

Seriously, unless you have been directly exposed to a patient with a verified case of 2019 NCoV, your risk of catching, being hospitalized, and dying from the flu is higher.

Again: You probably don’t need to be worried about dying from 2019 NCoV unless you have had contact with someone who had an active case of the disease. Statistically, you yourself (and the very young and very old that you care about) are at a much higher risk of catching the flu and dying from it than the Novel coronavirus.

You never get the flu? Good for you. Doesn’t mean you can’t catch it. You get sick after a flu shot? Not the shot’s fault (the shot contains no live/infectious viruses), you caught something that isn’t the flu. Sorry about your luck. Feeling a bit off the next day means that you are having a strong reaction to it.