Spiders (Arthropoda: Arachnida: Araneae) are fascinating and most diverse group of air breathing chelicerate arthropods in the animal kingdom. Their distribution is worldwide except Antartica and occupies every ecological niche available except of air and sea colonization. An improved air-dried method was to prepare spider chromosomes. Chromosome data are reported for 3 species belonging to families Pholcidae, Hersiliidae and Lycosidae from India. The number of diploid chromosomes (2n) in both males and females and sex determining mechanism of each species were determined as follows: Crossopriza lyoni: 2n=23 (XO ) male, Crossopriza lyoni: 2n=24 (XX ) female, Hersilia savignyi: 2n=30 (X 1 X 2 O) male, Hersilia savignyi: 2n=32 (X 1 X 2 X 3 X 4 ) female, Hippasa agelenoides: 2n=28 (X 1 X 2 O) male, Hippasa agelenoides: 2n=30 (X 1 X 2 X 3 X 4 ) female. During the first meiotic division 11, 14, 13 autosomal bivalents were found in Crossopriza lyoni, Hersilia savignyi and Hippasa agelenoides respectively. Crossopriza lyoni have metacentric and submetacentric type of chromosomes. In Hersilia savignyi chromosomes are acrocentric and rod shaped in Hippasa agelenoides.