With the matter coming to a head, Speaker P Dhanapal came out of the assembly escorted by marshals shortly after the House assembled to facilitate the floor test.
The proceedings were stalled amid the pandemonium, where some opposition members demanded secret voting.
Palaniswami, who was sworn in as Chief Minister two days ago, has opted to prove his majority today itself though he was given 15 days time by Governor Ch Vidyasagar Rao.
With the decision of the two, the Palaniswami camp is left with 122 MLAs in the 234-member House with one vacancy.
Ailing DMK supremo M Karunanidhi was not present in the House.
In another development, the Panneerselvam camp appointed Semmalai as the party whip in the assembly and sent a letter to the Speaker to this effect.